CN103443119B

CN103443119B - In amino acid/11 and 3 similar molecules of cyclosporin changed

Info

Publication number: CN103443119B
Application number: CN201180067449.4A
Authority: CN
Inventors: A.赫曼斯; B.W.芬斯克; D.J.特勒帕尼耶; M.D.阿贝尔; S.苏吉亚马; D.R.乌雷
Original assignee: Kang Zhuo Pharmaceutical Co
Current assignee: Hepion Pharmaceuticals Inc
Priority date: 2010-12-15
Filing date: 2011-12-14
Publication date: 2017-09-22
Anticipated expiration: 2031-12-14
Also published as: MY192981A; BR112013014890B1; CA2821009A1; JP6144627B2; SG10201510236XA; HK1243633A1; CN107496902A; KR20130124515A; KR102305423B1; EP2651965A4; US20140142033A1; JP2014505035A; EP2651965A1; DK2651965T3; MX2013006831A; PT2651965T; SI2651965T1; ES2705750T3; SG191069A1; RU2630690C2

Abstract

The invention discloses the Cyclosporin A analogues changed comprising amino acid/11 and 3 bit substituents according to lower formula (I).Disclosed compound includes including cyclophilin the compound that cyclophilin A has compatibility, and the compound reduces inhibitive ability of immunity with cyclosporin A and its only compared with 1 modified analog.

Description

In amino acid/11 and 3 similar molecules of cyclosporin changed

Technical field

The present invention relates to the molecule for the new analog for belonging to cyclosporin family, include the analog of cyclosporin A (CsA) With including it is with reduction or without immunosuppressive activity and combine cyclophilin (CyP) analog.

Background technology

Cyclosporin is the member of the ring type polypeptide species with effective immunosuppressive activity.At least partly this kind of chemical combination Thing (such as cyclosporin A (CsA)) be by species many spores wood category fungies (Tolypocladium inflatum) secondary metabolites Produce.CsA is effective immunodepressant, it has therefore proved that it can suppress humoral immunity and cell-mediated immune response, such as Allograft rejection, delayed allergy, experimental allergic encephalomyelitis, Freund's adjuvant arthritis and transplanting Thing versus-host disease.It is used in organ transplant prevent organ rejection；And for treating rheumatoid arthritis and use In treatment psoriasis.

Although many compounds are known in cyclosporin family, CsA is probably medically most widely used change Compound.CsA immunosuppressive effect is related to the activation event for suppressing T cell mediation.Immunosupress be by cyclosporin with it is general Store-through the intracellular protein that is referred to as cyclophilin (cyclophilin) (CyP) be combined and realize.The compound suppresses calcium again The calcium and the activity of calmodulin-dependence serine-threonine phosphatase of neural element enzyme.The suppression of calcium nerve element prevents transcription The factor such as NFAT_p/cWith NF- κ B activation, this for cytokine gene in t cell activation (IL-2, IFN-γ, IL-4 and GM-CSF) induction be required.

Since cyclosporin is originally found, huge variety of naturally occurring ring spore bacterium has been had isolated and identified Element.In addition, being prepared for many non-days by partially or completely synthesizing mean and by the cell culture technology of application improvement The cyclosporin so existed.Therefore, the species comprising cyclosporin consist essentially of for example naturally occurring cyclosporin A~Z； A variety of non-naturally occurring cyclosporin derivatives；Artificial or synthesis cyclosporin, including dihydro cyclosporin and different ring spore Rhzomorph；(3 '-O- atoms of such as MeBmt residues can be acylated derivative cyclosporin, or can be drawn on 3- flesh aminoacyl residues Enter other substituents)；Wherein there are cyclosporin (such as wherein MeBmt resi-dues 6 ' and 7 ' of isomeric form in MeBmt residues Being configured as on position is cis rather than trans)；The cyclosporin of variant amino acids is mixed wherein in peptide sequence on ad-hoc location.

In position 1, the cyclosporin analog containing modified amino acid is disclosed in WO 99/18120 and WO 03/033527 In, it is incorporated herein by reference in its entirety.These applications describe referred to as " ISA_TX247 " or " ISA247 " or " ISA " ring Spore streptozotocin derivative.The analog is identical with cyclosporin A in structure in addition to the modification on the residue of amino acid/11 position.Application People previously had found, some mixtures of ISA247 cis and trans isomers (including mainly include trans ISA247 mixing Thing) show enhancing inhibitive ability of immunity effect and reduce the compound action of toxicity, better than naturally occurring and ring spore that is being currently known Rhzomorph.

Have determined that cyclosporin there are three cell targets；Calcium nerve element, CyP hypotypes (including but not limited to CyP-A, CyP-B and CyP-D), and P- glycoprotein (PgP).The combination of cyclosporin and calcium nerve element causes significant immunosupress, and negative Duty and transplanting and the conventional contact of autoimmunity sign.

Cyclophilin family

CyPs (enzyme committee (EC) numbering 5.1.2.8) belongs to class protein, cis- trans with peptide acyl-prolyl Isomerase activity；This protein is referred to as immunophilin, and including FK-506- conjugated proteins and cellule albumen (parvulins).CyPs is found in all cells for all organisms (prokaryotes and eucaryote) studied, and In evolution guarded in structure.The mankind have 7 kinds of main CyPs, i.e. CyP-A, CyP-B, CyP-C, CyP-D, CyP-E, CyP- 40 and CyP-NK (identifies) from man day Natural killer cell first, amounts to 16 kinds of unique protein (Galat A. Peptidylprolyl cis/trans isomerases (immunophilins): biological diversity - targets - functions. Curr Top Med Chem 2003, 3:1315-1347；Waldmeier PC etc. .Cyclophilin D as a drug target. Curr Med Chem 2003, 10:1485-1506)。

The first CyPs member identified in mammal is CyP-A.CyP-A is 18-kDa cytosol albumen Matter, and be the most abundant protein that CsA is combined.It is estimated that total cell solute protein (the Mikol V of CyP-A compositions 0.6% Deng .X-ray structure of monmeric cyclophilin A-cycloporin A crystal complex at 2.1 A resolution. J. Mol. Biol. 1993, 234:1119-1130；Galat A, Metcalfe SM. Peptidylproline cis/trans isomerases. Prog. Biophys. Mol. Biol. 1995, 63:67- 118)。

The cell position of cyclophilin

CyPs can be had found in the most cells lacuna of most of tissues, and the unique function of coding.In mammal In, CyP-A and CyP-40 are cytosols, and CyP-B and CyP-C have the signal sequence of amino terminals, target it Endoplasmic reticulum albumen matter secretory pathway (is summarized in Galat, 2003；The .Structures of such as Dornan J immunophilins and their ligand complexes. Curr Top Med Chem 2003, 3:1392-1409).CyP-D has letter Number sequence, makes it point to mitochondria (the .Cyclophilins and their possible role such as Andreeva L in the stress response. Int J Exp Pathol 1999, 80:305-315；Hamilton GS etc. .Immunophilins: beyond immunosuppression. J Med Chem 1998, 41:5119-5143)；CyP- E has the RNA- binding domain of amino terminals, and positioned at karyon (the .A nuclear such as Mi H RNA-binding cyclophilin in human T cells. FEBS Lett 1996, 398:201-205), and CyP-40 has TPRs, And positioned at cytosol (.cyclophilin-40, a the protein with homology to the such as Kieffer LJ P59 component of the steroid receptor complex. Cloning of the cDNA and further characterization. J Biol Chem 1993, 268:12303-12310).People CyP-NK is maximum CyP, is had Huge hydrophily and positively charged c-terminus, and positioned at the cytosol (.A such as Anderson SK cyclophilin-related protein involved in the function of natural killer cells. Proc Natl Acad Sci USA 1993, 90:542-546；The The N-terminal such as Rinfret A cyclophilin-homologous domain of a 150-kilodalton tumor recognition molecule exhibits both peptidylprolyl cis-transisomerase and chaperone activities. Biochemistry 1994, 33:1668-1673)。

The function of cyclophilin and activity

CyPs is the multifunctional protein being related in many cell processes.Protected because CyPs is height all the time in evolution Keep, this shows that CyPs has vital effect.Initially, it has been found that CyPs has catalysis peptidyl-prolyl key along anteiso- Special enzyme performance (Galat, 1995 of structure；The .A phase I study of such as Fisher GA paclitaxel (taxol) (T) in combination with SDZ valspodar, a potent modulator of multidrug resistance (MDR). Anticancer Drugs.1994；5(Suppl 1): 43).Therefore, CyPs quilts Referred to as peptidyl-prolyl cis-trans isomerase (PPI enzymes), it can serve as acceleration in the protein newly synthesized is inherently folded The factor, PPI enzymes, which are also assisted in, to be repaired due to the protein that environmental stress is damaged, and the environmental stress includes heat stress, ultraviolet and shone Penetrate, the pH of cellular environment change and with the treatment of oxidant.This function is referred to as the molecular chaperoning activity (.Roles such as Yao Q of cyclophilins in Cancers and Other Organs Systems. World J. Surg. 2005, 29: 276-280)。

In addition, display recently, CyPs PPI enzymatic activitys are related to different cell processes, including intracellular protein transport (Andreeva, 1999；The .New member of the cyclophilin family such as Caroni P associated with the secretory pathway. J Biol Chem 1991, 266:10739-42), mitochondrial function (the .CsA binding to mitochondrial cyclophilin inhibits such as Halestrap AP the permeability transition pore and protects hearts from ischaemia/reperfusion injury. Mol Cell Biochem 1997, 174:167-72；Connern CP, Halestrap AP. Recruitment of mitochondrial cyclophilin to the mitochondrial inner membrane under conditions of oxidative stress that enhance the opening of a calcium- sensitive non-specific channel. Biochem J 1994, 302:321-4), mRNA precursor is processed (the .A serine/argininerich nuclear matrix cyclophilin such as Bourquin JP interacts with the Cterminal domain of RNA polymerase II. Nucleic Acids Res 1997, 25: 2055-61) and multiprotein complex Stability Maintenance (Andreeva, 1999).

Cyclosporin is in hydrophobic pocket via contact with nanomole affinity combination CyP-A (Colgan J etc. .Cyclophilin A-Deficient Mice Are Resistant to Immunosuppression by Cyclosporine. The Journal of Immunology 2005, 174:6030-6038, Mikol, 1993), and Suppress PPI enzymatic activitys.However, this effect is considered as incoherent with immunosupress.More correctly, between CsA and CyP-A Complex establish aggregate surface, it combines and prevents genetic transcription (the Friedman J of the plain regulating cell factor of calcium nerve Deng .Two cytoplasmic candidates for immunophilin action are revealed by affinity for a new cyclophilin: one in the presence and one in the absence of CsA. Cell 1991, 66: 799-806；The .Calcineurin is a common target such as Liu J of cyclophilin-CsA and FKBP-FK506 complexes. Cell 1991 , 66: 807-815)。

The homology of cyclophilin

CyP-A, typical family member is highly conserved protein (Handschumacher in mammalian cell The .Cyclophilin such as RE: a specific cytosolic binding protein for CsA.Science 1984, 226: 544-7).People CyP-A sequence homology analysis shows that it is very high homology with people CyP-B, CyP-C and CyP-D (Harding MW, Handschumacher RE, Speicher DW. Isolation and amino acid sequence of cyclophilin. J Biol Chem 1986, 261：8547-55).Pass through the pact in highly conserved region 109 amino acid, form all CyPs cyclosporin binding pocket.In known CyPs, CyP-D has with CyP-A's Highest homology.In fact, the sequence identity between CyP-A and CyP-D in this region is 100% (Waldmeier 2003；The .The Mitochondrial Permeability Transition as a Target such as Kristal BS for Neuroprotection. Journal of Bioenergetics and Biomembranes 2004, 36( 4)；309- 312).Therefore, CyP-A compatibilities are the extraordinary predictors of CyP-D compatibilities, the vice versa (.The such as Hansson MJ Nonimmunosuppressive Cyclosporine analogues NIM811 and UNIL025 Display Nanomolar Potencies on Permeability Transition in Brain-Derived Mitochondria.Journal of Bioenergetics and Biomembranes, 2004,36(4): 407-413).This relation is It is repeatedly available empirical proof (Hansson, 2004 of cyclosporin analog；The .Inhibition such as Ptak Rg of Human Immunodeficiency Virus Type 1 Replication in Human Cells by Debio-025, a Novel Cyclophilin Binding Agent.Antimicrobial Agents and Chemotherapy 2008: 1302-1317；The .Genetic and pharmacologic inhibition of such as Millay DP mitochondrial dependent necrosis attenuates muscular dystrophy. Nature Medicine 2008, 14 (4): 442-447；The .The Discovery of Novel such as Harris R Non-Immunosuppressive Cyclosporine Ethers and Thioethers With Potent HCV Activity. Poster # 1915,59th Annual Meeting of the American Association for the Study of Liver Diseases(AASLD), 2008).CyPs sequence homology proposes that all CyPs are the potential targets of cyclosporin analog Mark.Because numerous cell processes that CyPs is related to, this further proposes, keeps significantly can be used for controlling with reference to CyP CsA analogs Treat the idicatio of many diseases.

The disease of cyclophilin mediation

Human immunodeficiency virus (HIV):

HIV is the lentivirus of retrovirus family, and serves as CyP participations in some virus infection and reproduction process Example.CyP-A was just confirmed as effective target (Rosenwirth BA etc. in AntiHIV1 RT activity chemotherapy before 10 years .Cyclophilin A as a novel target in anti-HIV-1 chemotherapy. Int. Antivir. News1995, 3:62-63).CyP-A meets the basic function of early stage in HIV-1 replicative cycles.It is found specific knot Close the HIV-1 Gag polyproteins (Gag of the .Human immunodeficiency virus such as Luban JKL type 1 protein binds to cyclophilins A and B. Cell 1993, 73: 1067-1078).Capsid protein p24 (CA) amino acid sequence determined around G89 and P90 is accredited as CyP-A binding site (Bukovsky AAA etc. .Transfer of the HIV-1 cyclophilin-binding site to simian immunodeficiency virus from Macaca mulatta can confer both cyclosporine sensitivity and cyclosporine dependence. Proc. Natl. Acad. Sci.USA1997, 94: 10943-10948； The .Crystal structure of human cyclophilin A bound to the such as Gamble TRF amino- terminal domain of HIV-1 capsid. Cell1996, 87: 1285-1294).Affinity of the CyP-A to CA CyP-A is promoted to mix Virosome particles (the .Functional association such as Thali MA of in assembling cyclophilin A with HIV-1 virions. Nature1994, 372: 363-365).Experimental evidence shows, CyP-A-CA interaction is that HIV-1 duplications are essential；HIV-1 in human cell can be damaged by suppressing this interaction Duplication (the .Cyclophilin interactions with incoming such as Hatziioannou TD human immunodeficiency virus type 1 capsids with opposing effects on infectivity in human cells. J. Virol.2005, 79: 176-183；The .Mode of action such as Steinkasserer AR of SDZ NIM 811, a nonimmunosuppressive CsA analog with activity against human immunodeficiency virus type 1 (HIV-1): interference with early and late events in HIV-1 replication. J. Virol 1995, 69: 814-824).Answered in the CyP-A viruses being related to Step in cycle processed is proved to be the event before cellular genome is incorporated into double-stranded viruses DNA after virion permeates (the .Cyclophilin A is required for an early step in the life such as Braaten DEK cycle of human immunodeficiency virus type 1 before the initiation of reverse transcription. J. Virol 1996 70: 3551-3560；The .The such as Mlynar ED non- immunosuppressive CsA analogue SDZ NIM 811 inhibits cyclophilin A incorporation into virions and virus replication in human immunodeficiency virus type 1-infected primary and growth-arrested T cells. J. Gen. Virol 1996, 78: 825-835；Steinkasserer, 1995).The activity of CsA anti-HIV-1s is reported in 1988 first (the .The effect of CsA on infection of susceptible cells by such as Wainberg MA human immunodeficiency virus type 1. Blood 1998, 72: 1904-1910).CsA and many derivatives are pressed down The CsA analogs that the evaluation that HIV-1 processed is replicated discloses nonimmune suppression have the activity of anti-HIV-1, its equivalent to or very To better than those immunosuppressive analog (the .Inhibition of human such as Bartz SRE immunodeficiency virus replication by nonimmunosuppressive analogs of CsA. Proc. Natl. Acad. Sci.USA 1995, 92:The .Mode of action of SDZ such as 5381-5385, Billich AF NIM 811, a nonimmunosuppressive CsA analog with activity against human immunodeficiency virus (HIV) type 1: interference with HIV protein-cyclophilin A interactions.J. Virol 1995, 69: 2451-2461 ；Ptak, 2008).

Inflammation

Inflammation in disease is related to leucocyte (white blood corpuscle) and flows into infected zone.Leucocyte by chemotactic factor (CF), (inhale by chemistry Yin Ji families) it is attracted to the region.In vitro study shows that extracellular CyP-A is effective chemistry suction of human leukocytes and T cell Draw agent (the .Extracellular cyclophilins contribute to the such as Kamalpreet A regulation of inflammatory responses Journal of Immunology 2005；175: 517-522；Yurchenko The .Active-site residues of cyclophilin A are crucial for its such as VG signaling activity via CD147. J. Biol. Chem.2002；277: 22959-22965；The .Leukocyte such as Xu QMC chemotactic activity of cyclophilin. J. Biol. Chem.1992；267: 11968-11971； The .Interaction with glycosaminoglycans is required for cyclophilin such as Allain FC B to trigger integrin-mediated adhesion of peripheral blood T lymphocytes to extracellular matrix. Proc. Natl. Acad. Sci. USA 2002；99: 2714-2719).In addition, working as In vivo during injection, CyP-A, which can induce, rapidly is characterized in that inflammatory response (Sherry BN etc. that leucocyte is flowed into .Identification of cyclophilin as a proinflammatory secretory product of lipopolysaccharide-activated macrophages. Proc. Natl. Acad. Sci. USA1992；89: 3511-3515).CyP-A is generally distributed in the cell, but during inflammatory response, and CyP-A passes through living and dying thin Born of the same parents are discharged into ECT gap (Sherry, 1992).In fact, in several different diseases associated with inflammation (including sepsis Disease, rheumatoid arthritis) and vascular smooth muscle cells disease in, it has been reported that elevated levels of CyP-A (Jin ZG etc. .Cyclophilin A is a secreted growth factor induced by oxidative stress. Circ. Res.2000；87: 789-796；Teger, 1997；Billich, 1997).In the situation of rheumatoid arthritis, report Between CyP-A levels and neutrophil count in rheumatoid arthritis patients synovia positive correlation (Billich, 1997)。

Cancer

Recently in many cancerous tissues and cell line, include but is not limited in small and non-small cell lung cancer, carcinoma of urinary bladder, liver In cell cancer, cancer of pancreas and breast cancer, CyP-A shows overexpression (Li, 2006；The .Cyclophilin such as Yang H A is upregulated in small cell lung cancer and activates ERK1/2 signal.Biochemical and Biophysical Research Communications 2007；361：763-767；Campa, 2003).In the case where providing exogenous CyP-A, showing stimulates growth (Li, 2006 of cancer cell；Yang, 2007), and CsA prevents the growth (Campa, 2003).Recently, it has been demonstrated that CyP (A and B) participates in allowing human breast carcinoma intricately The biochemical route of cell growth, and CyP knock out growth, propagation and the motion (.The such as Fang F that experiment reduces cancer cell expression of Cyclophilin B is Associated with Malignant Progression and Regulation of Genes Implicated in the Pathogenesis of Breast Cancer. The American Journal of Pathology2009；174(1): 297-308；The .Prolyl such as Zheng J Isomerase Cyclophilin A Regulation of Janus-Activated Kinase 2 and the Progression of Human Breast Cancer. Cancer Research 2008；68 (19): 7769-7778).Most enjoyably, CsA is controlled The mouse of heterograft breast cancer cell is treated, neoplasm necrosis is induced and fully suppresses transfer (Zheng, 2008).Study people Member draws " cyclophilin B effect can significantly contribute to the pathogenesis of human breast carcinoma " and " suppression of cyclophilin is probably a kind of The new therapeutic strategy of human breast carcinoma treatment " (Fang, 2009；Zheng, 2008).

Hepatitis C

HCV (HCV) is most common liver diseases in the world, and epidemic disease is considered as by the World Health Organization.Cause For HCV can before being found infected patient many decades, it is commonly known as " tranquillization " epidemic disease.Research shows, full generation HCV, the total incidence of world population about 3.3% have been infected more than 200,000,000 people in boundary.Only in the U.S., nearly 4 million people infects or felt Contaminate HCV；Wherein 2,700,000 people undergo chronic infection.All HCV infection individuals are in the liver disease for just developing into serious life-threatening In the risk of disease.The Current standards treatment of chronic hepatitis C is two kinds of broad sense antivirotic Peg-IFN alpha-2bs and Li Bawei Combination (the .Clinical trial results of peginterferons such as the Craxi A in of woods combination combination with ribavirin. Semin Liver Dis 2003；23(Suppl 1): 35-46).This treatment Mortality be about 50% (Molino BF. Strategic Research Institute: 3^rd annual viral hepatitis in drug discovery and development world summit 2007. AMRI Technical Reports；12(1)).

Recently it has proven convenient that the key (.Cyclophilin such as the Watashi K B that CyP-B, which is HCV genomes, effectively to be replicated Is a Functional Regulator of Hepatitis C Virus RNA Polymerase.Molecular Cell 2005, 19: 111-122).Replicated for its efficient gene group, virus depends on the factor such as CyP-B of host derivation. CyP-B and HCV RNA polymerases NS5B interacts, directly to stimulate its RNA binding activity.RNA interference (RNAi) mediation The levels of replication for reducing HCV is lost caused by the reduction of endogenous CyP-B expression and NS5B combinations CyP-B.Therefore, CyP- B serves as the stimulation conditioning agent of NS5B in HCV duplicating mechanisms.This regulation mechanism to virus replication can differentiate that CyP-B is disease-resistant The target of malicious therapeutic strategy.

Unlike HCV other treatment, CyP suppresses not being directly targeted in HCV virus.Result, it is believed that to CyP combination medicines Slower (the The way forward in such as Manns MP HCV will occur obtaining than current HCV therapy medicine in tolerance treatment-finding the right path. Nature Reviews Drug Discovery 2007；6:991- 1000).In addition, by the interference in host-Virus Interaction level, CyP suppresses that the new way of HCV-Ab IgG treatment may be opened Footpath, it is not only to the treatment based on interferon, and to being directly targeted HCV replicase such as protease and AG14361 Future therapeutic can be complementary (Flisiak R, Dumont JM, Crabb é R. Cyclophilin inhibitors in hepatitis C viral infection. Expert Opinion on Investigational Drugs 2007, 16(9): 1345-1354).The exploitation of new anti-HCV medicament of HCV virus duplication is influenceed significantly by suitable for fear of lacking Laboratory HCV models.This obstacle is more recently by several suitable cell culture model (Subgenomic HCV of exploitation Replicon Systems) and mouse model (the Evaluation of the such as Goto K anti-containing human liver cell hepatitis C virus effects of cyclophilin inhibitors, CsA, and NIM811. Biochem Biophys Res Comm 2006；343: 879-884；The Hepatitis C virus such as Mercer DF replication in mice with chimeric human livers. Nat Med2001；7:927-933) just overcome.Ring spore bacterium Element has been proved anti-HCV activity (the CsA such as Watashi K suppresses in screening model and small clinical trials recently replication of hepatitis C virus genome in cultured hepatocytes. Hepatology 2003；38:1282-1288；Inoue K, Yoshiba M. Interferon combined with cyclosporine treatment as an effective countermeasure against hepatitis C virus recurrence in liver transplant patients with end-stage hepatitis C virus related disease. Transplant Proc 2005；37:1233-1234).

Muscle degeneration obstacle

CyP-D is the part of all cell Mitochondria Permeability Transition Pores (MTP).The function in MTP holes is to provide Intracellular calcium homeostasis.Under normal operation, the open and close of MTP holes is reversible.Be related to excessive calcium current enter it is intracellular Under pathological conditions, this makes the irreversible opening in mitochondria excess load and induction MPT holes, causes cell death or apoptosis.It is reported that It is mitochondrial that CsA correct for its in the patient of Jan Ullrich (Ullrich) congenital muscular dystrophy and Bethlam myopathies Dysfunction and muscle cell apoptosis [(the CsA corrects mitochondrial dysfunction such as Merlini L and muscle apoptosis in patients with collagen VI myopathies. PNAS 2008；105(13): 5225-5229].CsA has confirmed that it dose-dependently suppresses the mitochondrial MTP of isolated myocardium and opened in vitro, so as to hinder Only Apoptosis and permission cell have repair time (the Inhibition of such as the Gomez L mitochondrial of preciousness permeability transition improves functional recovery and reduces mortality following acute myocardial infarction in mice Am J Physiol Heart Circ Physiol 2007, 293: H1654-H1661).In there is the clinical research of patients of acute myocardial infarction at 58, and observed by placebo Result compare, it was confirmed that the Reperfu- sion time apply CsA (the Effect such as Piot C ofs related to less infarct Cyclosporin on Reperfusion Injury in Acute Myocardial Infarction. New England Journal of Medicine 2008；395(5): 474-481)).

Chronic neurodegenerative disease

CsA can be used as the neuroprotective agent (such as Keep M in acute cerebral ischemia and damaged condition due to head injury Intrathecal cyclosporine prolongs survival of late-stage ALS mice. Brain Research 2001；894: 327-331).It is relative there was only 10% survival rate when lacking treatment, show through the CsA animals treated Show noticeable 80% survival rate.Through determination later, this is mainly CsA combination mitochondrias CyP-D result.Through subsequent It is determined that, CsA effectiveness extends to chronic neurodegenerative change, as sick (ALS) by Lu Jialei (Lou Gerhig) family name later Rat model confirms (U.S. Patent number 5,972,924) that wherein CsA treatments add more than one times of residual life.Recently It is also shown that CyP-D inactivations protect experimental autoimmune encephalomyelitis (multiple sclerosis in CyP-D knock-out mices Animal model) aixs cylinder (the Cyclophilin D inactivation protects axons such as Forte M in experimental autoimmune encephalomyelitis, an animal model of multiple sclerosis. PNAS 2007；104(18): 7558-7563).In Alzheimer's mouse model, CyP-D lacks Substantially improve function (the Cyclophilin D deficiency such as the Du H attenuates of study, memory and cynapse mitochondrial and neuronal perturbation and ameliorates learning and memory in Alzheimer's disease Nature Medicine 2008, 14(10): 1097-1105).In addition, CsA is in henry It is proved to be effective (the CsA protects striatal such as Leventhal L neurons in the sick rat model in the court of a feudal ruler in vitro and in vivo from 3-nitropropionic acid toxicity. Journal of Comparative Neurology 2000, 425(4):471-478), and in parkinsonian mouse model it is partially effective (the CsA attenuates degeneration of dopaminergic neurons induced such as Matsuura K by 6-hydroxydopamine in the mouse brain. Brain Research 1996, 733(1): 101-104)。 Therefore, mitochondrio-dependant necrosis represents the pathogenesis of protrusion, it is proposed that the new pharmacology of these diseases can be provided by suppressing CyP-D Learn therapeutic strategy (Du, 2008).

Due to metabolic defect in cellular calcium ion (Ca ²⁺ ) damage of cell, tissue and organ lost of stable state

Ca²⁺Many physiological processes are participated on a cellular level, include the mitochondrial function of health.In some pathological conditions such as In myocardial infarction, apoplexy, acute liver toxicity, storage/reperfusion injury of cholestasis and transplant organ, mitochondria, which is lost, to be adjusted The ability of calcium level is saved, excessive calcium accumulation causes the substantial amounts of hole of mitochondrial inner membrane to open (Rasola A in mitochondrial matrix Deng The mitochondrial permeability transition pore and its involvement in cell death and in disease pathogenesis. Apoptosis 2007, 12: 815-833).Pass through hole The up to non-selective conduction of the ion of 1.5 kilodaltons and molecule, is referred to as the process of mitochondrial permeability transformation, causes line Plastochondria expands and reaches that cell death includes other events of inducing cell apoptosis.One of MTP composition is CyP-D.CyP-D It is immunophilin molecule, its isomerase activity adjusts MPTP opening, suppressing isomerase activity by CsA or CsA analogs can press down MPTP processed is produced, so as to prevent cell death.

The cyclosporin analog cyclophilin inhibitor of nonimmune suppression

Although favourable effects of the CsA in above-mentioned idicatio, the effect that immunosupress occurs together limits CsA as CyP The practicality of inhibitor in clinical practice.At present, only minority CsA analogs have been demonstrated the immune suppression seldom or reduced System activity is (i.e.<10% CsA immunosupress efficiency), and still retain its with reference to CyP ability (i.e. compared with CsA>10% knot Conjunction ability).

NIM 811(Melle ⁴ - cyclosporin)

NIM 811 be the snow-white curved neck of fungi it is mould (Tolypocladium niveum) tunning, it is in 4, amino acid On changed and show no immunosuppressive activity (being combined due to lacking calcium nerve element), but retain to CyP-A binding affinity (the Inhibition of human immunodeficiency virus such as Rosenwirth BA type 1 replication by SDZ NIM 811, a nonimmunosuppressive Cyclosporine Analogue.Antimicrob Agents Chemother 1994, 38: 1763-1772)。

DEBIO 025 (MeAla ³ EtVal ⁴ - cyclosporin)

DEBIO 025 is the CsA of the double base chemical modification on amino acid 3 and 4.DEBIO 025 is displayed that and is not immunized Inhibitory activity, still also retains the binding affinity (Kristal, 2004) to CyP-A PPI enzymatic activitys.

SCY-635 (the thio Sar of dimethylaminoethyl ³ - hydroxyl Leu ⁴ - cyclosporin)

The CsA of SCY-635 double base chemical modifications on amino acid 3 and 4.SCY-635 displays that no immunosupress is lived Property, still also retain the binding affinity (PCT Publication WO2006/039668) to CyP-A PPI enzymatic activitys.

In general, these compounds have the modification to being responsible for combining in terms of the CsA of calcium nerve element, and generally require and repair Change amino acid 3 and 4.Amino acid 3 and the modification of 4 are arduous and complicated, because this way is usually directed to open loop spore The ring of rhzomorph, replace and/or change these amino acid and then close ring to produce the cyclosporin of modification.

In contrast, the modification of side chain amino acid 1 need not open the ring of cyclosporin.However, amino acid/11 position with CyP combines (rather than combining calcium nerve element) correlation, and modification adds CsA immunosuppressive effect.Such as U.S. Patent number 6,605,593 disclose the single modification of amino acid/11 position, and it causes CsA analogs to have increased immunosupress efficiency.

Therefore, it is being readily synthesized and the effective similar molecule of cyclosporin (" CAM ") should in the disease for the treatment of CyP mediations Make us desired to possess.Also make us desirably providing CsA analogs, the analog provides at least some CsA original function, But it has property improve or extra, effect and function relative to original CsA.

Summary of the invention

According to one aspect, compound of the invention is similar including the cyclosporin A according to nonimmune suppression defined herein Thing.According on the other hand, the compounds of this invention includes cyclophilin A to cyclophilin has compatibility.According to other side, the present invention Compound include Cyclosporin A analogues, its disease mediated to cyclophilin or illness are useful, and have developed these diseases The therapy of disease or illness.

According on one side, the present invention relates to formula L compound：

Wherein

A. R' is H or acetyl group；

B. R1 is the saturation of 2 to 15 carbon atom lengths or the aliphatic carbon chain of undersaturated straight or branched；

C. R2 is selected from：

i. H；

Ii. unsubstituted, N- substitutions or N, N- disubstituded amide；

Iii. the amine that the substituted or unsubstituted acyl groups of N- are protected；

Iv. carboxylic acid；

V. the substituted or unsubstituted amine of N-；

Vi. nitrile；

Vii. ester；

Viii. ketone；

Ix. the alkyl of hydroxyl, dihydroxy, trihydroxy or polyhydroxy；With

X. substituted or unsubstituted aryl；

Xi. the aliphatic chain of saturation or undersaturated straight or branched, it optionally includes and is selected from hydrogen, ketone, hydroxyl, nitrile, carboxylic Acid, ester, 1,3- dioxolanes, the substituent of halogen and oxo；

Xii. aromatic group, it includes the substituent selected from halide, ester and nitro；With

Xiii. the combination of the aliphatic chain (xi) and aromatic group (xii) of saturation or undersaturated straight or branched； With

D. R23 is the optionally substituted aliphatic carbon chain of saturation or undersaturated straight or branched.

In one aspect, substituent R 1-R2 is selected from：

。

In one aspect, R2 is selected from：

Wherein

I. R5 is the saturation of 1 to 10 carbon lengths or the aliphatic carbon chain of undersaturated straight or branched；With

Ii. R6 is monohydroxylated, dihydroxy, the saturation of trihydroxy or polyhydroxylated 1 to 10 carbon lengths or not The aliphatic carbon chain of the straight or branched of saturation.

In one aspect, saturations of the substituent R 1-R2 comprising 2 to 5 carbon or the aliphatic of undersaturated straight or branched Chain, it is optionally selected from the substituent substitution of hydrogen, ketone, hydroxyl, nitrile, halogen, oxo, carboxylic acid, ester and 1,3- dioxolane；

In one aspect, R3 is selected from：

On the one hand, R23 includes optionally substituted alkyl, including optionally substituted C1-C3 alkyl.The alkyl can be with Replaced by amino, and C1-C3-Ala can be included, wherein the compound includes D- epimers.In described implementation In scheme, R23 can be MeAla.

On the one hand, above-mentioned formula L

Selected from following：

With

。

On the one hand, R23 is the fat of the straight or branched of 1~6,1~5,1~4,1~3 or 2 carbon length Fat race carbochain.

On the one hand, the present invention relates to the method for treating or preventing the disease that cyclophilin is mediated in mammal, it is included in Under conditions of the disease or injury for the treatment of cyclophilin mediation compound as described herein, or the compound are applied to mammal Or the purposes of disease or injury described in composition treatment, or the compound is in the medicine that preparation is used for the purposes or treatment Purposes.The disease or injury can be overexpressed to mediate by cyclophilin, or the disease is the congenital overexpression of cyclophilin 's.The disease or injury of the cyclophilin mediation can be selected from：

A. virus infection；

B. diseases associated with inflammation；

C. cancer；

D. muscular disorders (muscular disorder)；

E. neurological disorder；With

F. lost with ischemic, Reperfu- sion, cell calcium homeostasis, ionic homeostasis is lost, free radical production increases or induction line grain The related damage of the toxin of body function obstacle；

Wherein described virus infection is optionally by selected from caused by following virus：Human immunodeficiency virus, A type liver Inflammation, hepatitis B, hepatitis C, hepatitis D, Hepatitis E, SARS-CoV, hCoV-NL63, hCoV-HKU-1, hCoV- OC43, hCOV-229E, coronavirus, feline infectious peritonitis virus and infectious gastroenteritis virus (transmissible gastroenteritis virus)；

Wherein described diseases associated with inflammation is optionally selected from asthma, autoimmune disease, chronic inflammation, chronic prostate Inflammation, glomerulonephritis, anaphylactia, inflammatory bowel disease, septicemia, vascular smooth muscle cells disease, aneurysm, pelvic inflammatory disease Disease, reperfusion injury, rheumatoid arthritis, graft rejection and vasculitis；

Wherein described cancer be optionally selected from small and non-small cell lung cancer, carcinoma of urinary bladder, hepatocellular carcinoma, cancer of pancreas, breast cancer, Glioblastoma, colorectal cancer, squamous cell carcinoma, melanoma and prostate cancer；

Wherein described muscular disorders are optionally selected from myocardial reperfusion injury, DMD, collagen VI myopathies Syndrome (PCAS), heart failure, atherosclerosis and abdomen master after (collagen VI myopathies), heart arrest Aneurysm；

Wherein described neurological disorder is optionally selected from Alzheimer's, Parkinson's disease, Huntington's disease, multisystem Atrophy, multiple sclerosis, cerebral paralysis, epilepsy, apoplexy, diabetic neuropathy, ALS (Lu Jialeishi Disease), bipolar disorders, exitotoxicity damage (excitotoxic injury), hepatic encephalopathy, hypoglycemia, manganese poisoning (manganese toxicity), neuron target deprive (neuronal target deprivation), toxicity aliphatic acid such as Arachidonic acid, mechanical nerve injury, spinal cord injury and brain damage；With

The wherein described damage related to the forfeiture of cell calcium homeostasis be optionally selected from myocardial infarction, apoplexy, acute liver toxicity, Storage/reperfusion injury of cholestasis and transplant organ.

On the one hand, the present invention relates to the method for preparing above-mentioned formula L compounds, it comprises the following steps：

1) cyclosporin A (CsA) is deposited with alkaline alkyl lithium amide (lithium alkylamide) in suitable solvent Reacted lower, then with the reaction of suitable electrophilic reagent with the compound of production 1：

2) by the compound of formula 1 and AC₂O is reacted in the presence of suitable solvent, to form formula 2A compound：

3) formula 2A compound and oxidant are reacted, to form formula 3A compound：

4) formula 3A compound and electrophilic reagent are reacted, to form formula 4A compound：

5) optionally formula 4A compound is carried out deacetylated.

On the one hand, above-mentioned formula L preparation, which is included in the solvent, adds excessive LiCl to primarily form formula L L- Epimer, or the preparation of the formula L are carried out under LiCl missings, to primarily form formula L D- epimers.The alkali Property alkyl amino lithium may include lithium diisopropylamine.

On the one hand, the electrophilic reagent is selected from the group defined in following table, to generate the corresponding R23 proposed in table：

On the one hand, the present invention relates to the method for preparing formula L defined herein, it comprises the following steps：

1) formula 1A compound is reacted with dimethylamino naphthyridine in the presence of appropriate solvent, to form formula 2 Compound：

Optionally subsequently form the aldehyde of formula 3：

Optionally then by Wittig reaction with production L compound, wherein R23 is selected from：

On the one hand, the present invention relates to formula L defined herein preparation, it comprises the following steps：By the compound of formula 5 with Alkaline alkyl lithium amide, optionally including lithium diisopropylamine, in the presence of suitable electrophilic reagent, in appropriate solvent Reacted, to form formula L compound, wherein R23 includes optionally substituted C1-C3 alkyl.

For all chemical formulas disclosed in this document：

" carboxylic acid " includes the group that wherein carboxylic moiety is connected to one of following substituent：

1. the alkyl (alkyl of such as 2 to 15 carbon) that can be substituted；

2. the alkenyl (alkenyl of such as 2 to 15 carbon) that can be substituted；With

3. the alkynyl (alkynyl of such as 2 to 15 carbon) that can be substituted；

Above-mentioned substitution may include halogen (such as fluorine, chlorine, bromine, iodine), nitro, cyano group, hydroxyl, the sulfydryl that can be substituted (such as sulfydryl (thiol), C1-4 alkyl sulfenyl (alkylthio)), the amino that can be substituted (such as amino, single-C1-4 Alkyl amino, two-C1-4 alkyl aminos, 5- to 6- member ring type amidogen for example nafoxidine, piperazine, piperidines, morpholine, thiomorpholine, Pyrroles, imidazoles etc.), can by halo C1-4 alkoxies (for example methoxyl group, ethyoxyl, propoxyl group, butoxy, trifluoromethoxy, Trifluoro ethoxy etc.), can by halo C1-4 alkoxy -C 1-4 alkoxies (for example methoxymethoxy, methoxy ethoxy, Ethoxy ethoxy, trifluoromethoxy ethyoxyl, trifluoroethoxy base oxethyl etc.), formoxyl, C2-4 alkanoyls (such as acetyl Base, propiono etc.) and C1-4 alkyl sulphonyls (such as mesyl, ethylsulfonyl) etc., and the number of substitution is preferably 1 To 3.

In addition, the substitution of above-mentioned " amino that can be substituted " can be combined with each other to form ring type amidogen (for example, group is from 5- The upper removing hydrogen of nitrogen atom ring such as nafoxidine, piperazine, piperidines, morpholine, thiomorpholine, pyrroles, imidazoles etc. to 6- yuan of rings is former Son, so that substitution may be connected to what is formed on nitrogen-atoms etc.).Ring type amidogen group can be substituted, and the example bag of substitution Include halogen (such as fluorine, chlorine, bromine, iodine), nitro, cyano group, hydroxyl, sulfydryl (such as sulfydryl, C1-4 alkyl that can be substituted Sulfenyl etc.), the amino that can be substituted (such as amino, single-C_1-4Alkyl amino, two-C1-4 alkyl aminos, the ring ammonia of 5- to 6- members Base such as nafoxidine, piperazine, piperidines, morpholine, thiomorpholine, pyrroles, imidazoles etc.), can be esterified or amidated carboxyl (for example Carboxyl, C1-4 alkoxy-carbonyls, carbamoyl, list-C1-4 alkyl-carbamoyls, two-C1-4 alkyl-carbamoyls Deng), can be by C1-4 alkoxies (such as methoxyl group, ethyoxyl, propoxyl group, butoxy, trifluoromethoxy, the trifluoroethoxy of halo Base etc.), can be by the C1-4 alkoxy -Cs of halo_1-4Alkoxy (such as methoxymethoxy, methoxy ethoxy, ethoxy ethoxy Base, trifluoromethoxy ethyoxyl, trifluoroethoxy base oxethyl etc.), formoxyl, C2-4 alkanoyls (such as acetyl group, propiono Deng), C1-4 alkyl sulphonyls (such as mesyl, ethylsulfonyl), and the number of substitution is preferably 1 to 3.

" amine " includes group, and it is unsubstituted or wherein amine moiety is that N- substitutions or N, N are dibasic, and with one It is individual or two can be independently selected from following substituent：

1. the alkyl (alkyl of such as 2 to 15 carbon) that can be substituted；

2. the alkenyl (alkenyl of such as 2 to 15 carbon) that can be substituted；

3. the alkynyl (alkynyl of such as 2 to 15 carbon) that can be substituted；

4. formoxyl or acyl group (alkanoyl (such as acetyl group, propiono, the butyryl of such as 2 to 4 carbon that can be substituted Base, isobutyryl etc.) and 1 to 4 carbon alkyl sulphonyl (such as mesyl, ethylsulfonyl) etc.)；

5. the aryl that can be substituted (such as phenyl, naphthyl)；Etc.；

And be connected on substituent, the substituent is independently selected from as above substituent defined in " carboxylic acid ".

" acid amides " includes compound, and the carboxylic group of wherein amide moieties, which is connected to, to be independently selected from as above " carboxylic acid " and defined On the substituent of substituent, the amino group of connection amide moieties is that N- replaces or N, N are dibasic, and has one respectively Or two can be independently selected from following substituent：

1. the alkyl (alkyl of such as 2 to 15 carbon) that can be substituted；

2. the alkenyl (alkenyl of such as 2 to 15 carbon) that can be substituted；

3. the alkynyl (alkynyl of such as 2 to 15 carbon) that can be substituted；

5. the aryl that can be substituted (such as phenyl, naphthyl)；Etc.

" aryl " can be by monocyclic or fused polycycle aromatic hydrocarbon group for example, for example preferred C6-14 aromatic yl groups such as benzene Base, naphthyl, anthryl, phenanthryl or acenaphthenyl (acenaphthylenyl) etc., preferably phenyl.The aryl can be taken by one or more For base substitution, the substituent such as lower alkoxy (such as C1-6 alkoxies as methoxyl group, ethyoxyl or propoxyl group), halogen Atom (such as fluorine, chlorine, bromine, iodine), low alkyl group (such as C1-6 alkyl as methyl, ethyl or propyl group), low-grade alkenyl (such as C2-6 alkenyls as vinyl or pi-allyl), low-grade alkynyl (such as C.2-6 alkynyl as acetenyl or propargyl), The amino that can be substituted, the hydroxyl that can be substituted, cyano group, the amidino groups that can be substituted, carboxyl, elementary alkoxy carbonyl (such as C1- 6 alkoxy carbonyls such as methoxycarbonyl or ethoxy carbonyl etc.), the carbamoyl that can be substituted (for example can be by C1-6 alkyl Or acyl group (for example formoxyl, C2-6 alkanoyls (alkanoyl), benzoyl, can by the C1-6 alkoxy carbonyls of halo, can C1-6 alkyl sulphonyls, benzenesulfonyl by halo etc.) substitution carbamoyl, the alkyl or acyl group can by 5- to 6- member Aromatic monocyclic heterocyclic group (such as pyridine radicals), 1- azelidinyls carbonyl, 1- pyrrolidinylcarbonyls, piperidino carbonyl Base, morpholino carbonyl, thiomorpholine replace for carbonyl (sulphur atom can be oxidized), 1- piperazinyl carbonyls etc.), etc..It is any this A little substituents can independently be replaced 1 to 3 alternative position.

The carbonyl group of " ketone " including wherein ketone part is connected to the compound on one or two substituent, the substitution Base is independently selected from substituent defined in " carboxylic acid " as described above.

" ester " includes carboxylate or alcohol ester, and wherein ester group includes one or two substituent, and the substituent is independently selected From substituent defined in " carboxylic acid " or " aryl ".

" alkyl " unless otherwise defined, the alkyl of preferably 1 to 15 carbosilane unit length.

" aromatic group " can be illustrated as aryl as defined above, or 5- to 6- members aromatic monocyclic heterocyclic group, Such as furyl, thienyl, pyrrole radicals, oxazolyl, isoxazolyls, thiazolyl, isothiazolyl, imidazole radicals, pyrazolyl, 1,2,3- Evil Di azoly, 1,2,4- oxadiazolyls, 1,3,4- oxadiazolyls, furan a word used for translation base (furazanyl), 1,2,3- thiadiazolyl groups, 1,2,4- Thiadiazolyl group, 1,3,4- thiadiazolyl groups, 1,2,3- triazolyls, 1,2,4- triazolyls, tetrazole radical, pyridine radicals, pyridazinyl, pyrimidine Base, pyrazinyl, triazine radical etc.；With the heterocyclic group of (preferably 10- to 12- members) aromatics fusion of 8- to 16- members.

" nonimmune to suppress " refers to the ability that compound suppression normal human lymphocyte proliferation is measured in cell culture and preferred During the method measurement proposed in example 1 below 9, the compound shows that substantially reduction suppresses immune system compared with CsA The ability of level.

" analog " refers to CsA analogue, and it is different from CsA in one or more functional groups.Preferably, so Analog at least retain the critical parts of CsA combination CyP abilities.

The preferred kind of Formulas I is：Wherein R' is H, and R1 is the saturation or undersaturated alkyl of 2 to 15 carbon lengths, and R2 It is selected from：

1. the carboxylic carboxylic acid of bag；

2. the N of N- substitutions, N- disubstituded amides, wherein substituent independently selected from H, the alkyl of 1 to 7 carbon lengths, Or the substituent formation heterocycle, wherein heterocycle is selected from O, N or S；

3. the ester of 1 to 7 carbon lengths；

4. monohydroxylated or dihydroxy the alkyl of 1 to 7 carbon lengths；

5. the amine of 1 to 7 carbon lengths of the substituted or unsubstituted acyl group protections of N-；

6. nitrile；

7. the carboxyl of ketone, wherein ketone is connected to R1, and saturation or the length of undersaturated alkyl chain are 1 to 7 carbon；

8. the phenyl being optionally substituted by one or more substituents, the substituent be independently selected from nitrogen dioxide, fluorine, amine, Ester or carboxyl.

The compound of the present invention can exist in the form of optically active compound.The present invention includes optics in the range of above formula All enantiomers of reactive compound, each individual enantiomer and racemic mixture.Equally, the present invention includes defined hereinization The pro-drug of compound.

According on the other hand, the compounds of this invention can be used for the CyP for treating or preventing or studying the preferred people of mammal to be situated between The disease led.This disease is typically to be overexpressed mediation by CyP, such as the congenital overexpression mediations of CyP.

The disease for the CyP mediations that can be treated by the compounds of this invention includes：

A. virus infection；

B. diseases associated with inflammation；

C. cancer；

D. muscle degeneration obstacle (muscular degenerative disorder)；

E. neurodegenerative disorders (neurodegenerative disorder)；With

F. the damage related to the forfeiture of cell calcium homeostasis.

It is described virus infection be probably by selected from human immunodeficiency virus, hepatitis A, hepatitis B, hepatitis C, Caused by the virus of hepatitis D and Hepatitis E.The diseases associated with inflammation is selected from asthma, autoimmune disease, chronic inflammation Disease, chronic prostatitis, glomerulonephritis, anaphylactia, inflammatory bowel disease, septicemia, vascular smooth muscle cells disease, artery Knurl, pelvic inflammatory disease, reperfusion injury, rheumatoid arthritis, graft rejection and vasculitis.The cancer is optional from childhood With non-small cell lung cancer, carcinoma of urinary bladder, hepatocellular carcinoma, cancer of pancreas and breast cancer.The muscle degeneration obstacle may be selected from cardiac muscle again Perfusion injury, DMD and collagen VI myopathies.The neurodegenerative disorders may be selected from Alzheimer's, handkerchief gold Sen Shi diseases, Huntington's disease, multi-system atrophy, multiple sclerosis, cerebral paralysis, apoplexy, diabetic neuropathy, amyotrophia Lateral sclerosis (Lu Jialei diseases), spinal cord injury and brain damage.The damage related to the forfeiture of cell calcium homeostasis may be selected from cardiac muscle Infarct, apoplexy, acute liver toxicity, storage/reperfusion injury of cholestasis and transplant organ.

Detailed description

According to one aspect, compound of the invention can be applied to warm-blooded animal in need merely or together with pharmaceutical carriers With.Pharmaceutical carriers can be solid or liquid.Compound can with containing Conventional nontoxic pharmaceutically acceptable carrier, adjuvant, With the preparation of the unit dose of excipient, by oral administration, topical application, it is parenteral, suction spraying or rectally apply.This paper institutes Include hypodermic injection, intravenous, intramuscular, breastbone inner injection or perfusion technique with term is " parenteral ".

Pharmaceutical composition containing inventive mixture can be the form for being adapted to oral application, such as tablet, lozenge, ingot Agent, aqueous or oily suspensions, dispersible powder or particle, emulsion, hard or soft capsule or syrup or elixir.It is intended to oral The composition of application can be prepared according to the method for preparation pharmaceutical composition known in the art, and such composition can contain one Plant or a variety of medicaments selected from sweetener, flavouring, colouring agent and preservative, to provide pharmaceutically graceful and tasty preparation. Containing the tablet with the active component that acceptable excipient is mixed on non-toxic pharmaceutical, it can also be prepared by known method. Excipient used can be for example：(1) inert diluent, such as calcium carbonate, lactose, calcium phosphate or sodium phosphate；(2) pelletize and be disintegrated Agent, such as cornstarch or alginic acid；(3) adhesive, such as starch, gelatin or Arabic gum；(4) lubricant, such as stearic acid Magnesium, stearic acid or talcum.Tablet can be uncoated or they can be with known technology coatings, so as in delaying stomach and intestine road Disintegration and absorb, so as to provide lasting effect in longer in the period of.For example, material such as glycerine list can be postponed with application time Stearate or glycerol distearate.They can also use U.S. Patent number 4,256,108；4,160,452；With 4,265,874 Described in technology coatings, to form the osmotic therapeutic tablets of control release.

In some cases, formulations for oral can be hard gelatin capsule forms, and wherein active component is consolidated with inertia Body diluent such as calcium carbonate, calcium phosphate or kaolin mixing.They can also be soft gelatin capsules, wherein active component Mixed with water or oil medium such as peanut oil, atoleine or olive oil.

Waterborne suspension is usually contained with being adapted to prepare the active component that the excipient of waterborne suspension is mixed.It is such to assign Shape agent may include：(1) it is suspending agent, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methyl cellulose, sodium alginate, poly- Vinylpyrrolidone, bassora gum and Arabic gum；Or (2) dispersant or wetting agent, it can be naturally occurring phosphatide such as ovum The contracting of the condensation product of phosphatide, alkylene oxide and aliphatic acid such as Myrj 45, oxirane and long-chain fatty alcohol Product such as heptadecaethylene oxycetanol (heptadecaethyleneoxycetanol), oxirane and partial ester is closed (to be derived from Aliphatic acid and hexitol) condensation product such as polyoxyethylene 80 sorbitan monooleate or oxirane and partial ester (be derived from fat Fat acid and hexitan) condensation product such as SPAN 80.

Waterborne suspension can also contain one or more preservatives, such as ethyl-para-hydroxybenzoate or P-hydroxybenzoic acid N-propyl；One or more colouring agents；One or more flavourings；With one or more sweeteners, such as sucrose, aspartame or Saccharin.

Oily suspensions can be by being suspended in vegetable oil (such as peanut oil, olive oil, sesame oil or coconut by active component Oil), be made by formula in the fish oil containing omega-3 fatty acid or mineral oil (such as atoleine).Oily suspensions can contain thickening Agent, such as beeswax, hard paraffin or hexadecanol.Also sweetener and flavouring can be added, to provide tasty oral formulations.These Composition can be preserved by adding antioxidant such as ascorbic acid.

Dispersible pulvis and granule are suitable for preparing waterborne suspension.They provide with dispersant or wetting agent, Suspending agent and the active component of one or more preservative mixing.Suitable dispersant or wetting agent and suspending agent by it is above-mentioned that It is a little to illustrate.Also other excipient may be present, for example, the above sweetener, flavouring and colouring agent.

Pharmaceutical composition containing inventive mixture can also be oil-in-water emulsion form.Oil phase can be vegetable oil (such as olive oil or peanut oil) or mineral oil (such as atoleine) or their mixture.Suitable emulsifying agent can be (1) Naturally occurring natural gum, such as Arabic gum and bassora gum；(2) naturally occurring phosphatide such as soybean and lecithin；(3) it is derived from fat The ester or partial ester 30 of fat acid and hexitan, such as dehydrating sorbitol monooleate；(4) condensation of the partial ester and oxirane Product, such as SPAN 80.Emulsion can also contain sweetener and flavouring.

Syrup and elixir can together be prepared with sweetener (such as glycerine, propane diols, sorbierite, aspartame or sucrose). Such preparation can also contain moderator (demulcent), preservative and flavouring and colouring agent.

Pharmaceutical composition can be aqueous or oily suspensions the form of Sterile injectable.The suspension can be according to known Those suitable dispersants or wetting agent and suspending agent that method application has already mentioned above are prepared.Sterile injectable preparation also may be used With in being the aseptic parenteral solution or suspension in nontoxic parenteral acceptable diluent or solvent, such as 1,3-BDO Solution.Wherein available acceptable solvent and solvent are water, Ringer's solution and isotonic sodium chlorrde solution.In addition, sterilizing Fixed oil is conventionally used as solvent or suspension media.For this purpose, any gentle fixed oil can be applied, including The monoglyceride or diglyceride of synthesis.In addition, aliphatic acid such as oleic acid can be applied in ejection preparation preparation.

The compounds of this invention can also be administered with suppository form, the rectally for the medicine.Suitable composition can By by the compound be at normal temperatures solid but under rectal temperature be liquid appropriate nonirritant excipient mix and It is made, and therefore can melts to discharge medicine in the rectum.Such material is cocoa butter and polyethylene glycol.

For topical application, can be used the appropriate usual creme applied together with cyclosporin, ointment, gel, solution, Or suspension etc..

In particularly preferred embodiments, using containing surfactant, ethanol, lipophilic and/or amphiprotic solvent conduct The liquid solution of non-active ingredient.Especially, using oral containing this isomeric analogue mixture and following non-medical ingredients Multiple emulsion is formulated：D- alpha tocopherol cetomacrogol 1000 succinates (vitamin E TPGS), medium chain triglyceride (MCT) oil, Tween 40 and ethanol.Also preferably application contains the compound and the soft gelatin glue with oral administration solution identical non-medical ingredients Capsule (includes gelatin, glycerine, water and D-sorbite).

It should be appreciated, however, that many factors will be depended on for the specific dosage level of any particular patient, including it is used The activity of particular compound, the age, body weight, general health, sex, diet, administration time, method of administration, drainage rate, Compatibility of drugs and the specified disease or the property of illness and seriousness just treated.

Methodology

Reaction proposed below is on the amino acid/11 position residue (AA1) and 3 residues (AA3) of amino acid for can synthesize CsA The general example of the chemical reaction of the required compound of modification.AA1 modification is described as follows：

Modification with AA3 is described as follows：

AA1 and AA3 modifications all should use the reagent with required chemical property, and it will be understood by those skilled in the art that can make The replacement of some reactants.

The identification of prepared compound and purity are generally obtained by the methodology including mass spectrography, HPLC and NMR spectra It is determined that.Mass spectrum (ESI-MS) is determined in the MSD systems of Hewlett Packard 1100.NMR spectra is in Varian On the MHz spectrometers of MercuryPlus 400 deuterated solvent (phosphonium salts be DMSO, every other compound be benzene) in carry out Determine.Analysis and the reversed-phase HPLC prepared are carried out on the serial systems of Agilent 1100.

PhosphoniumThe synthesis of salt compound

Phosphonium salt passes through triphenylphosphine or any other suitable phosphine and alkyl halide (R-X；X=Cl, Br or I) reaction It is made.Suitable alkyl halide is any uncle or any secondary aliphatic halide of any chain length or molecular weight.These Alkyl halide can for branch or non-branch, saturation or undersaturated alkyl halide.

If reaction is carried out (reaction 1) in toluene, the product directly Precipitation from reaction solution.However, non-live Property the more polar solvents of substrate requirements such as dimethylformamide (DMF) (reaction 2), to shorten the reaction time and reach satisfied Yield.

Reaction 1:

Wherein X is halide (including but is not limited to Cl, Br and I), and R10 is saturation or undersaturated straight or branched Aliphatic chain, its optionally include selected from ketone, hydroxyl, nitrile, carboxylic acid, ester and 1,3- dioxolane substituent；Aromatic group, It optionally includes the substituent selected from halide, ester and nitro；Or above-mentioned saturation or the aliphatic of undersaturated straight or branched The combination of chain and above-mentioned aromatic group.

The 404-15 of embodiment 1. synthesis

As illustrative example, triphenylphosphine (13 mmol) is dissolved in 50mL toluene, chlroacetone (10 is added Mmol) to obtain clear solution.Reaction is stirred overnight under reflux.Colorless solid is filtered out, washed with toluene and hexane, very Sky is dried.

Using reaction 1, following compounds are the compounds for the more embodiments that can be synthesized.

Alternative, Shi Dang phosphonium salts can be synthesized by reaction 2 as follows：

Reaction 2:

The 404-51 of embodiment 2. synthesis

As illustrative example, triphenylphosphine (11 mmol) is dissolved in 10 mL DMF, 4- bromo-butyric acids (10 are added mmol).Reaction is stirred 7 hours at 110 DEG C, is then allowed to cool overnight.50mL toluene is added, crystallization is collected by filtration Colorless solid.The product is washed with toluene and hexane, is dried in vacuum overnight.

If not starting to crystallization after being handled with toluene, by product with 20 mL MeOH/H₂O (1 :1 mixing Thing) extraction.Aqueous phase is washed with toluene and hexane, and is dried.Residue is being flowed back together with 50 mL ethyl acetate (EtOAc) At a temperature of stir 20-30 minutes.If obtaining crystalline solid, product is collected by filtration, washed with EtOAc and hexane, Dry.In the case where obtaining product for oil or colloid, EtOAc is decanted, by remaining product in vacuum drying.

Using reaction 2, following compounds are the compounds for the more embodiments that can be synthesized.

Wittig reacts

Wittig (Wittig) reacts the substrate and reactant for being widely used in wide scope.The side of substrate is introduced in reaction Chain, can represent any number of branch and non-branch, saturation and unsaturated variable-length aliphatic compound (R'), and The functional group of wide scope can be included.

In Wittig reaction, alkali such as potassium tert-butoxide (KOtBu) produces inner salt for You phosphonium salts.Inner salt and substrate Carbonyl (CsA- aldehyde) reacts, to generate alkene.The alkali of at least two equivalents is needed to produce inner salt containing carboxylic acid side chain phosphonium salts.

Reaction 3：Acetylation cyclosporin analog intermediate reacts the synthesis of Shi phosphonium salt compounds by Wittig

Wherein X is halide (including but is not limited to Cl, Br and I), and R12 is saturation or undersaturated straight or branched Aliphatic chain, its optionally include selected from ketone, hydroxyl, nitrile, carboxylic acid, ester and 1,3- dioxolane substituent；Aromatic group, It optionally includes the substituent selected from halide, ester and nitro；Or above-mentioned saturation or undersaturated, straight or branched aliphatic The combination of chain and above-mentioned aromatic group.

The compound 404-20 of embodiment 3. reacts the synthesis of Shi phosphonium salt compounds by Wittig：

As illustrative example, 250 mL flasks of drying are loaded into butyl triphenyl phosphonium bromide under an argon atmosphere (6.0 mmol) and 40 mL anhydrous tetrahydro furans (THF).Suspension is cooled to 0 DEG C, potassium tert-butoxide (6.0 mmol) is added, It is orange to obtain.Reaction is stirred 1-2 hours in environment temperature, being subsequently added CsA- aldehyde, (2.0 mmol are dissolved in 20 mL anhydrous THF).Continue to be stirred at room temperature 3 hours.React with 10 mL saturations NH₄Cl and 20 mL ice water quenchings.Each layer is separated, aqueous phase is used EtOAc is extracted.Organic layer is merged, salt water washing is used, through Na₂SO₄Dry.Remove solvent, and by crude product through silica gel (hexane/ Acetone 3:1) purify.

Using reaction 3, following compounds are the compounds for the more embodiments that can be synthesized.

It is deacetylated

Reaction 4：Acetylation cyclosporin analog it is deacetylated

Wherein R12 is the aliphatic chain of saturation or undersaturated straight or branched, its optionally include selected from ketone, hydroxyl, nitrile, Amine and the substituent of 1,3- dioxolanes that carboxylic acid, ester, acid amides, acyl group are protected；Aromatic group, it optionally includes and is selected from halogenation Thing, ester, the substituent of amine and nitro；Or the aliphatic chain and above-mentioned aromatic series base of above-mentioned saturation or undersaturated straight or branched The combination of group.

Embodiment 4：Compound 404-90 is through deacetylated synthesis

As illustrative example, by solution of the 404-20 (0.16 mmol) in 10 mL MeOH and tetramethyl hydrogen Amine-oxides pentahydrate (0.47 mmol) is in 2 mL H₂Solution in O merges.Mixture is stirred at room temperature 2 days.In vacuum Concentration reaction, adds 5 mL H₂O water.Use EtOAc extractive reactions, extract salt water washing, through Na₂SO₄Dry, be concentrated into It is dry.The HPLC purifying of the inverted preparation of crude product.

The purifying of deacetylated compound typically passes through silica gel (hexane/acetone 2:1) or by the HPLC of preparation carry out.Changing In the case of compound 404-60,404-137,416-08,420-98 and 420-100 (carboxylic acid), 1 M is used into reaction before extraction HCl is acidified to pH value for 2-3.

Using reaction 4, following compounds are the compounds for the more embodiments that can be synthesized.

The hydrogenation of double bond

Double bond can be hydrogenated under atmospheric pressure, to obtain the side chain of saturation.Functional group such as hydroxyl, carbonyl and carboxyl are at these It is stable under part, it is not required that protection.R' represents acetyl group or hydrogen.In the case of α, beta-unsaturated carbonyl compound, Double bond must be reduced before deacetylation, to avoid cyclisation of the free hydroxyl to activity double key nucleophilic addition.

Reaction 5：

Wherein R12 is the aliphatic chain of saturation or undersaturated straight or branched, its optionally include selected from ketone, hydroxyl, nitrile, Amine and the substituent of 1,3- dioxolanes that carboxylic acid, ester, acid amides, acyl group are protected；Aromatic group, it optionally includes and is selected from halogenation Thing, ester, the substituent of amine and nitro；Or above-mentioned saturation or undersaturated, straight or branched aliphatic chain and above-mentioned aromatic series The combination of group, and R' is H or acetyl group.

Embodiment 5：404-56 synthesis

As illustrative example, 404-43 (0.34 mmol) is dissolved in the anhydrous EtOH of 40 mL, 43 mg Pd/ are added C (10 %) and 0.2 mL acetic acid.Stirred the mixture under hydrogen atmospheric pressure 2 days.Reaction is filtered by diatomite, vacuum Concentration.HPLC purifying of the crude product through preparation.

Using reaction 5, following compounds are the compounds for the more embodiments that can be synthesized.

The reduction of itrile group

Itrile group is reduced to corresponding primary amine can be by sodium borohydride (NaBH₄) and nickel chloride (II) (NiCl₂) generate in the original location Nickel borides is completed.Suitable trapping reagent is added, the primary amine (being respectively carbamate or acid amides) of acyl group protection is formed, and Prevent from forming the secondary amine as side reaction is not intended to.Double bond partial reduction under the specified conditions, and obtain product mixture.It is full It is separated and purifies with undersaturated shielded aminated compounds.For reaction 420-123, mixture is not separated by. On the contrary, by mixture through catalytic hydrogenation, to produce fully saturated compound.

Reaction 6：

Wherein acyl group is any one of BOC, acetyl group or bytyry, and acylating agent is di-tert-butyl dicarbonate, acetic acid Any one of acid anhydride and butyric anhydride, and the aliphatic group that R1 is saturation or undersaturated straight or branched.People in the art Member is it should be appreciated that above-mentioned acylating agent can be substituted by broad range of acylating agent, to produce same broad range of acyl group protection Amine.

Embodiment 6：420-08 synthesis

As illustrative example, 404-187 (0.257 mmol) is dissolved in 15 mL methanol, and be cooled to 0 DEG C.Plus Enter di-tert-butyl dicarbonate (0.514 mmol) and nickel chloride (II) (0.025 mmol), to obtain clear solution.Through 1 small time-division Part adds sodium borohydride (3.85 mmol).Resulting mixture is stirred overnight at ambient temperature.In 0 DEG C of addition Other sodium borohydride (1.95 mmol), and continue stirring 3 hours in room temperature.HPLC is shown as 420-08-1 (carbamates Class compound) and 420-08-2 (carbamate compound that double bond is reduced) mixture.By the reaction and divinyl Triamine (0.257 mmol) is stirred 30 minutes together, is then being concentrated in vacuo.Residue is absorbed with 75 mL EtOAc, uses 20 mL Saturation NaHCO₃Solution is washed, and through Na₂SO₄Dry.Solvent is removed in a vacuum.HPLC purifying of the crude product through preparation.

Using reaction 6, following compounds are the compounds for the more embodiments that can be synthesized.

The deprotection of amine

Trifluoroacetic acid (TFA) can be used to be converted into free amine through acidic hydrolysis for the amine (carbamates) of BOC protections.

Reaction 7：

Wherein R1 is saturation or undersaturated, straight or branched aliphatic chain, and R' is H or acetyl group.

Embodiment 7：420-23 synthesis

As illustrative example, 420-17 (0.026 mmol) is dissolved in the anhydrous DCM of 4 mL, and 2 are added at 0 DEG C ML trifluoroacetic acids.Reaction is stirred at room temperature 3 hours.Add 20 mL dichloromethane.Reactant mixture H₂O and saturation NaHCO₃Solution is washed, through Na₂SO₄Dry.Remove solvent, HPLC purifying of the crude product through preparation.

Using reaction 7, following compounds are the compounds for the more embodiments that can be synthesized.

The protection of amino group

The method that broad range of blocking group application can be used to set up for free amino function is protected.With from nitrile The reduction of beginning is introduced and compared, and more broad range of protective agent is available.In a word, reaction 7 and 8 provides the replacement of reaction 6 Route, the amino-compound for preparing acyl group protection.

Reaction 8：

Wherein acyl group is any one of BOC, acetyl group or bytyry, and acylating agent is di-tert-butyl dicarbonate, acetic acid Any one of acid anhydride and butyric anhydride, it will be appreciated by those skilled in the art that including the broad range of of two carbonic esters, acid anhydride and carboxylic acid halides Acylating agent can be used for the amine for producing broad range of acyl group protection, and R1 is saturation or the aliphatic of undersaturated straight or branched Group.

Embodiment 8：420-27 synthesis

As illustrative example, 420-25 (0.039 mmol) is dissolved in 3 mL anhydrous pyridines under a nitrogen.Will be anti- 0 DEG C should be cooled to, acetic anhydride (0.59 mmol) is added.Mixture is stirred overnight at ambient temperature.Solvent is being removed in vacuum, Residue is absorbed with 25 mL EtOAc.Reaction the M HCl of 2 x, 10 mL 1, the mL saturations NaHCO of 2 x 10₃Solution and 10 ML salt water washings, and through Na₂SO₄Dry.Solvent is removed in a vacuum, obtains the product of colorless solid.

The deprotection of aldehyde

1,3- dioxolanes part is converted into aldehyde functional group by acidic hydrolysis.

Reaction 9 and embodiment 9：404-47 synthesis

As illustrative example, solution of the 404-33 (0.246 mmol) in 20 mL formic acid is stirred at room temperature 45 minutes.100mL frozen water and 200 mL saturations NaHCO are slowly added into reaction₃Solution (strong foaming).React with 2 x 150 ML EtOAc are extracted.By the saturation NaHCO of the extract after merging₃Solution, water and salt water washing, and through Na₂SO₄Dry.True It is aerial to remove solvent and drying.

The reduction of nitryl group

Aromatic nitro compound is aniline by catalytic hydrogen reduction.The reaction causes the reduction of double bond.

Reaction 10 and embodiment 10：404-120 synthesis

As illustrative example, 404-89 (0.13 mmol) is dissolved in 2 mL ethanol, blue Buddhist nun (Raney) is added Nickel (0.18 g, in 50% H₂In O, washed 3 times, be then suspended in 2 mL ethanol with ethanol) and 0.1 mL acetic acid.In room temperature Stirring reaction 2 days.Reaction is filtered by diatomite, and filter cake is washed with methanol.Filtrate is dried.Residue is absorbed with EtOAc, is used NaHCO₃Solution and salt water washing, through Na₂SO₄Dry.Solvent is removed in vacuum.Crude product is through silica gel (hexane/acetone 2:1) it is pure Change.

The synthesis of acid amides

Acid amides is made (reaction 11) by amine with the reaction of corresponding acid chloride by carboxylic acid.The synthesis can also pass through application Appropriate coupling reagent such as DCC and HOBt are directly carried out (reaction 12) by sour.

Reaction 11：

Wherein R1 is saturation or undersaturated, straight or branched aliphatic chain, R15 and R16 stand alone as hydrogen or saturation or Undersaturated, straight or branched aliphatic chain, or wherein NR15R16 form morpholine base section together.

Embodiment 11：404-85 synthesis

As illustrative example, by 365-73 (0.04 mmol) and thionyl chloride (68 mmol) in a nitrogen atmosphere Merge, and be heated to reflux 2 hours.Allow reaction to cool down, and be concentrated to dryness.20mL toluene is added, reaction is concentrated to dryness (2 repeatedly It is secondary).Residue is absorbed with 5 mL dry toluenes, adds diethylamine (0.48 mmol).Reaction is stirred at room temperature overnight.Add 5mL H₂O, mixture is extracted with 20 mL EtOAc.By extract salt water washing, through Na₂SO₄Dry.Solvent is removed in vacuum, slightly Product is through silica gel (hexane/acetone 3:1) purify.

Using reaction 11, following compounds are the compounds for the more embodiments that can be synthesized.

Reaction 12：

Embodiment 12：420-104 synthesis

As illustrative example, 420-98 (0.078 mmol) is dissolved in the anhydrous DCM of 10 mL in a nitrogen atmosphere. Dicyclohexylcarbodiimide (DCC, 0.117 mmol) and I-hydroxybenzotriazole hydrate (HOBt, 0.078 are added at 0 DEG C Mmol), stir the mixture for 15 minutes.Dimethylamine (0.78 mmol) is added, the solution of clear colorless is obtained.Removed after 15 minutes Cooling bath is gone, continues to stir 5 days at ambient temperature.Reaction is transferred in separatory funnel, 20 mL DCM and 10 mL are added 0.5 M HCl.Organic layer is removed, through Na₂SO₄Dry, be concentrated to dryness.Residue is absorbed with 10 mL acetonitriles.Insoluble is consolidated Body is filtered out, and filtrate is concentrated in a vacuum.HPLC purifying of the crude product through preparation.

Using reaction 12, following compounds are the compounds for the more embodiments that can be synthesized.

Esterification

Using acidic catalyst (reaction 13) or coupling reagent, (14) DCC and DMAP, reaction prepared by corresponding carboxylic acid and alcohol Carboxylate.

Reaction 13：

Wherein R1 is saturation or undersaturated, straight or branched aliphatic chain, and R17 is saturation or undersaturated, straight The aliphatic chain of chain or side chain, the substituent optionally comprising halogen or hydroxyl.

Embodiment 13：404-171 synthesis

As illustrative example, by 404-60 (0.059 mmol), 4 mL EtOH and the 2 dense H of μ L₂SO₄Mixture It is heated to reflux 4 hours.Evaporation solvent, residue is absorbed with acetonitrile.HPLC purifying of the crude product through preparation.

Using reaction 13, following compounds are the compounds for the more embodiments that can be synthesized.

Reaction 14：

Embodiment 14: 420-24

As illustrative example, 404-60 (0.053 mmol) is dissolved in the anhydrous DCM of 4 mL in a nitrogen atmosphere, And it is cooled to 0 DEG C.Add dimethylamino naphthyridine (DMAP, 0.005 mmol), 2- fluorine propyl alcohol (0.27 mmol) and dicyclohexyl Carbodiimide (DCC, 0.058 mmol), and stir reaction 15 minutes at 0 DEG C.Cooling bath is removed, is continued at ambient temperature It is stirred overnight.20 mL DCM are added, then by reaction H₂O is washed, and is evaporated to dryness.Residue is absorbed with 10 mL acetonitriles, mistake Filter.It is being concentrated in vacuo filtrate.HPLC purifying of the crude product through preparation.

Alcohol

In addition to directly being synthesized in being reacted in Wittig, alcohol can be obtained by many reactions.Carbonyl with sodium borohydride also Original, forms primary alconol (by aldehyde) or secondary alcohol respectively (since ketone).

By the method for hydroboration, the oxidation of double bond can form the mixture of isomer.Reaction is main with anti- Geneva (anti-Markovnikov) direction is carried out.In the case of terminal olefine, primary alconol is major product.

Alkene is convertible into dihydric alcohol by the oxidation of hydrogen peroxide.Carbonyls and grignard reagent reacting, are formed secondary Alcohol (by aldehyde) and the tertiary alcohol (by ketone).This method allows carbochain to extend.

Reaction 15:

Wherein R' is H or acetyl group, and R1 is saturation or undersaturated, straight or branched aliphatic chain, and R20 is saturation Or undersaturated, straight or branched aliphatic chain.

Embodiment 15：404-98 synthesis

As illustrative example, 404-61 (0.0365 mmol) is dissolved in 4.5 mL in a nitrogen atmosphere anhydrous EtOH.Sodium borohydride (0.15 mmol is suspended in the anhydrous EtOH of 0.5 mL) is added at 0 DEG C, by resulting mixture It is stirred overnight at ambient temperature.Other sodium borohydride (0.08 mmol) is added, and continues to be stirred overnight.The reaction is in ice Bath cooling is lower to be quenched with 5 mL 1M HCl, is extracted with EtOAc.Extract salt water washing, through Na₂SO₄Dry, and be concentrated into It is dry.HPLC purifying of the crude product through preparation.

Using reaction 15, following compounds are the compounds for the more embodiments that can be synthesized.

Reaction 16:

Wherein R1 is saturation or undersaturated, straight or branched aliphatic chain.

Embodiment 16:420-28-1 synthesis

As illustrative example, 404-16 (0.081 mmol) is dissolved in the anhydrous THF of 4 mL in a nitrogen atmosphere.Will Reaction is cooled to 0 DEG C, adds BH₃THF (1M THF solution, 0.06 mmol).The reaction is stirred at room temperature overnight.HPLC tables Bright reaction is imperfect.Add other BH₃THF (0.5 mmol), continues to be stirred at room temperature 4 hours.Reaction is cooled to 0 DEG C, add 1.0 mL 1M NaOH and the % hydrogenperoxide steam generators of 0.30 mL 30.The mixture is stirred at room temperature overnight.With 25 ML EtOAc extract the reaction.Extract salt water washing, through Na₂SO₄Dry, and be concentrated to dryness.The product is through preparation HPLC is purified.

Reaction 17:

Wherein R1 is saturation or undersaturated, straight or branched aliphatic chain, and R' is H or Acetyl Groups.

Embodiment 17:420-49 synthesis

As illustrative example, 420-49 (0.037 mmol) is dissolved in the anhydrous THF of 5 mL under an argon atmosphere.Will Reaction is cooled to -70 DEG C, adds allylmgcl (1 M THF solution, 0.22 mmol).Reaction stirs 15 points at -70 DEG C Clock, then allows it to return to room temperature.After 90 minutes, reaction saturation NH₄Cl solution is quenched.Being extracted with 25 mL EtOAc should Reaction.Extract salt water washing, through Na₂SO₄Dry, and be concentrated to dryness.HPLC purifying of the product through preparation.Obtain acetylation With the mixture of deacetylated compound.

Reaction 18:

Wherein R1 is saturation or undersaturated, straight or branched aliphatic chain, and R23 is saturation or undersaturated, straight The aliphatic chain of chain or side chain.

Embodiment 18:404-126 synthesis

As illustrative example, 404-16 (0.054 mmol) is dissolved in 1 mL formic acid, hydrogen peroxide (30 % are added The aqueous solution, 0.52 mmol).The reaction is stirred at room temperature overnight, and is then concentrated to dryness.25mL EtOAc are dissolved the residue in, are used Saturation NaHCO₃Solution is washed, through Na₂SO₄Dry.Solvent is removed in a vacuum.Reaction 9 mL THF and 3 mL 1M NaOH Absorb, be stirred at room temperature 4 hours.Solvent is removed, residue is allowed in 25 mL EtOAc and 5 mL H₂Distributed between O.Organic layer Salt water washing is used, through Na₂SO₄Dry.Evaporation solvent, HPLC purifying of the crude product through preparation.

Embodiment 19:The modification that amino acid is 3

The substitution on AA3 that CsA experience is outlined below.Reacted with excessive LDA (lithium diisopropylamine), cause difference Four azepine enol lithium (lithium azaenolate) units and lithium alkoxide (lithium are included on the side chain of amino acid/11 position Alkoxide) unit and on AA3 (lithium enolate) enol lithium unit six lithium derivative (hexalithio derivative).With the substitution product on subsequent reactions generation AA3 (methyl amimoacetic acid) residue of appropriate electrophile.Suitable parent Electric reagent is such as alkyl halide, aldehyde, carbon dioxide and alkyl disulfide (table 1).Depending on reaction condition, it can obtain relative Two kinds of epimers of D and L of ratio.Approach A (seeing below) mainly results in D products, and approach B (LiCl of excessive addition) is obtained To the mixture of two kinds of epimers.

Embodiment 19:Substitution reaction on the AA3 of cyclosporin A.Obtain D and L stereoisomers.

Approach A：[D-MeSar]³-CsA

Under an argon atmosphere, the anhydrous THF of 160 mL and diisopropylamine (2.07 mL, 14.8 will be added in drying flask mmol).Solution is cooled to -78 DEG C, n-BuLi (2.5 M hexane solutions, 5.4 mL, 13.5 mmol) is added.30 points of stirring Zhong Hou, adds CsA (2.40 g, 2.0 mmol are dissolved in the anhydrous THF of 40 mL).Reaction is stirred 1 hour at -78 DEG C.Add another Outer n-BuLi (3.2 mL, 8.0 mmol), is subsequently added into methyl iodide (1.25 mL, 20.0 mmol).In -78 DEG C of continuation Stirring 1.5 hours, then allows reaction to be warmed to room temperature within 1.5 hours through other.Add 20 mL H₂0, THF is removed in a vacuum. Add 50 other mL H₂0, extracted with 150 mL EtOAc.By extract salt water washing, through Na₂SO₄Dry.In vacuum Middle removing solvent, crude on silica gel (hexane/acetone 3:1) purify.Yield：0.74 g (0.61 mmol, 30 %).

Approach B:[MeSar]³-CsA

Under an argon atmosphere, the anhydrous THF of 7.5 mL and diisopropylamine (0.46 mL, 3.3 will be added in 100 mL dry combustion methods bottle mmol).Solution is cooled to 0 DEG C, n-BuLi (1.32 mL, 2.5 M hexane solution, 3.3 mmol) is added.Reaction is 0 DEG C stirring 20 minutes, be subsequently cooled to -78 DEG C.CsA (601 mg, 0.5 mmol) and lithium chloride (636 are prepared under an argon Mg, 15 mmol) solution in the anhydrous THF of 12 mL and it is cooled to -78 DEG C.Then LDA solution is transferred to by this by sleeve pipe In mixture.Reaction is stirred 2 hours at -78 DEG C.Other n-BuLi (1.20 mL, 3.0 mmol) is added, is subsequently added into Methyl iodide (0.62 mL, 10 mmol).Mixture is risen to -20 DEG C, and stirred 3 hours at this temperature.Reaction is allowed to rise to room Temperature, uses saturation NH₄Cl solution is quenched, and is extracted with EtOAc (mL of 2 x 20), salt water washing is used, through Na₂SO₄Dry.In a vacuum Remove solvent, crude on silica gel (hexane/acetone 3:1) purify.Yield：[L-MeAla³]-CsA: 302 mg (0.25 mmol, 50 %)。[D-MeAla³]-CsA: 76 mg (0.06 mmol, 12 %)。

Table 1：The embodiment of available electrophilic reagent is alkylated on cyclosporin 3- positions.

The embodiment 20 and 21 being listed below is the general example of chemical reaction, and the chemical reaction, which can be used, possesses necessity The tube- nursery CsA of chemical property amino acid/11 and the required compound of 3 modifications, and it will be understood by those skilled in the art that can To carry out the displacement of some reactants.

Embodiment 20：It is alkylated CsA AA1 modifications

Embodiment 20 is provided changes the synthetic route that the preceding 3- positions in CsA introduce substituent in AA1 side chains.In 3- alkane After base, the aldehyde (compound 3 in the examples below) of the program formation acetylation of 2 steps, it is via Wittig reaction The suitable substrates of 1- modifications.This method allows introduced residue to AA1 side chains, AA1 side chains used in step 1~3 such as highly basic With under the reaction condition of oxidant have limited stability.

More case summaries of the compound prepared using the order are in table 2.

Step 1：The alkylation of AA3 side chains

As described above, being synthesized respectively according to approach A or B.

Step 2：Acetylating hydroxyl groups on AA1 side chains

[D-MeSar] will be added in the flask of drying under a nitrogen³- CsA (1.84 g, 1.51 mmol), N, N- diformazans Base aminopyridine (19 mg, 0.15 mmol) and 20 mL anhydrous pyridines, then add acetic anhydride (10 mL, 0.1 mol).Instead It should be stirred overnight at ambient temperature.Mixture is poured into 100 mL frozen water, stirred until all ice-outs.Solid passes through It is collected by filtration, dries in atmosphere.The solid is dissolved in 50 mL EtOAc, with 1M HCl (2x), saturation NaHCO₃Solution and salt Water washing.Organic phase is through Na₂SO₄Dry, evaporation.Crude on silica gel (hexane/EtOAc/MeOH 10:10:0.5) purify.

Step 3：The formation of aldehyde

10 mL dioxanes and 10 mL H are added into the flask containing compound 2 (800 mg, 0.636 mmol)₂0。 Add Nal0₄(544 mg, 2.54 mmol) and Os0₄(7.9 mM solution, 1：In 1 water/dioxanes, 4.05 mL, 32 Mmol), reaction is stirred at room temperature to stay overnight.Add 75 mL H₂0, with the mL EtOAc extractive reactions of 3 x 25.Extract water, Saturation NaHCO₃Solution, water and salt solution (each 25 mL) washing, through MgSO₄Dry.Solvent, crude on silica gel are removed in a vacuum (hexane/EtOAc 3:1) purify.

Step 4：Wittig reaction

Triphenyl -6- caproic acids phosphonium bromide (90 mg, 0.195 mmol) and 5 will be added in the flask of drying under an argon The anhydrous THF of mL.Potassium tert-butoxide (1M THF solution, 0.39 mL, 0.39 mmol) is added at 0 DEG C, solution is stirred 30 points Clock, obtains bright orange.Compound 3 is added dropwise into reaction, and (81 mg, 0.065 mmol are dissolved in 1 mL anhydrous THF), continue to be stirred at room temperature overnight.Use saturation NH₄Reaction is quenched in Cl solution, is extracted with EtOAc.By extract salt solution Washing, through Na₂S0₄Dry.Solvent, crude on silica gel (toluene/acetone 3 is being removed in vacuum:1) purify.

Step 5:It is deacetylated

Compound 4 (30 mg, 0.022 mmol) is dissolved in 2 mL methanol and 0.5 mL water, tetramethylammonium hydroxide five is added Hydrate (12 mg, 0.066 mmol).Reaction a couple of days is stirred at room temperature, until HPLC confirms that deprotection is completed.1M is used in reaction It is 2 that HCl, which is acidified to pH, is concentrated in a vacuum.Residue is dissolved in EtOAc, is washed with water, through Na₂S0₄Dry.Boil off molten Agent, HPLC purifying prepared by crude product.

The schematic diagram of the cyclosporine derivative of 1,3- modifications.

Table 2：Using the method for embodiment 20, following compounds are compound (X and the Y ginsengs for the more embodiments that can be synthesized It is admitted to and states diagram；The R referred in X represents the attachment structure with CsA AA1).

The alkylation of AA1 modified compounds

Reaction 21 introduces substituent on the AA3 residues of previous AA1 modified side chains compound.Except 19 available bases of reaction Group is outer, and the approach allows to be unstable in the case where AA3 introduces substituent, substituent reaction condition used in reaction 20 , such as, may experience oxidation in aldehyde forming process of sulphomethyl (thiomethyl) residue in this method step 3.

Embodiment 21

Under an argon atmosphere, the anhydrous THF of 1.5 mL and diisopropylamine (87 μ L, 0.62 will be added in 25 mL dry combustion methods bottle mmol).Solution is cooled to 0 DEG C, n-BuLi (2.5 M, in hexane, 0.25 mL, 0.62 mmol) is added.Mixture Stirred 20 minutes at 0 DEG C, be subsequently cooled to -70 DEG C.- 70 DEG C by clear and bright LDA solution be transferred to 404-76 (118 mg, 0.095 mmol) and lithium chloride (120 mg, 2.84 mmol) in the anhydrous THF of 1.5 mL solution.Continue to stir at -70 DEG C 2 hours.Other n-BuLi (0.23 mL, 0.58 mmol) is added, methyl iodide (118 μ L, 1.89 are subsequently added into mmol).Allow reaction to be warming up to -20 DEG C, and keep staying overnight at this temperature.Use saturation NH₄Reaction is quenched in Cl solution, uses EtOAc Extraction.Extract salt water washing, through Na₂S0₄Dry, and be evaporated to dryness.Crude on silica gel (hexane/acetone 3: 1→ 2: 1) purify.

Table 3：(X and Y are according to Fig. 3 for the embodiment of the compound prepared by method 21；The R referred in X represents the AA1 with CsA Attachment structure).

¹Isomers is not separated；² m⁺Signal.

The other modification of AA1 (or AA3, respectively) residue functional group can be carried out, to obtain a variety of derivative compounds, such as Ester, acid amides, alcohol etc..By the way that the double bond set up in Wittig reaction is reduced, saturated compounds can be obtained.

Embodiment 22：Acid amides formation-the 440-08 of carboxylic acid synthesis

In a nitrogen atmosphere, 440-02 (48 mg, 0.037 mmol) is dissolved in the anhydrous DCM of 5 mL, and is cooled to 0 DEG C. Add dicyclohexylcarbodiimide (DCC, 1 1.6 mg, 0.056 mmol) and I-hydroxybenzotriazole (HOBt, 5.0 mg, 0.037 mmol), mixture is stirred 15 minutes at 0 DEG C.Add dimethylamine (2 M THF solution, 0.19 mL, 0.38 Mmol), and in room temperature continue to stir 3 days.Reaction is diluted with 20 mL DCM, is washed with the M HCl of 15mL 0.5.Organic phase is passed through Na₂S0₄Dry, then dry to dry.HPLC purifying of the crude mixture through preparation.

Embodiment 23：Ester formation -440-31 synthesis

440-20 (30 mg, 0.022 mmol) is dissolved in the anhydrous EtOH of 4 mL and the 2 dense H of μ L₂S0₄.Reaction is heated back Stream 3 hours, is then allowed to cool to room temperature.Dry reaction is to dry.HPLC purifying of the crude product through preparation.

Embodiment 24：Acid amides formation (anti-acid amides) -440-15 of nitrile compound synthesis

Under a nitrogen, 440-09 (80 mg, 0.061 mmol) and 5 mL MeOH will be added in 50 mL flask.Instead 0 DEG C should be cooled to, Ni (II) Cl is added₂-6H₂0 (1.4 mg, 0.006 mmol) and acetic anhydride (19 μ L, 0.20 mmol). 2 batches of 2 hours intervals are divided to add sodium borohydrides (104 mg, 2.75 mmol).Then reaction is allowed to be warmed to room temperature and be stirred overnight.Instead After the completion of answering, the M HCl of 7 mL 1 are added.Concentrate solution is only about half of to its original volume in a vacuum.Gained mixture is used EtOAc is extracted, extract saturation NaHCO₃Solution and salt water washing, through Na₂S0₄Dry.Solvent is removed in a vacuum.The production Thing, it includes some saturated compounds, is not further purified in following steps.

Embodiment 25：440-25 synthesis

440-15 (83 mg, 0.061 mmol) is dissolved in the anhydrous EtOH of 10 mL.Addition palladium (on 10 wt %, carbon, 8 ) and 3-4 drop acetic acid mg.Reaction carries out hydrogenation a couple of days at room temperature and atmospheric pressure, until confirming that reaction is complete by HPLC.Pass through Diatomite filtering reaction, filtrate is evaporated to dryness.Crude product is purified by the HPLC of preparation, then carries out deacetylation step.

Embodiment 26：440-32 synthesis

440-25 (41 mg, 0.03 mmol) is dissolved in 4mL MeOH, tetramethylammonium hydroxide pentahydrate (16 is added Mg, 0.09 mmol, are dissolved in 1 mL H₂0).Reaction is stirred at room temperature 2 days.Concentration reaction in a vacuum.Add 5 mL H₂0, Product is extracted with EtOAc.By extract salt water washing, through Na₂S0₄Dry, and be evaporated to dryness.HPLC prepared by crude product Purifying.

Table 4：By the way that the double bond set up in Wittig reaction is reduced into the cyclosporine compounds that obtained 1,3- is modified Derivative embodiment (X and Y are according to Fig. 3；The attachment structure with CsA AA1 is represented with the R referred in X).

The suppression of cyclophilin A isomerase is determined

By 1, the 3 CsA analogs of the present invention, scheme and slightly changed according to scientific literature, enzymatic determination is for surveying Measure the suppression of CyP-A activity.The measure is catalyzed the peptide containing proline from cis to the conformation of transisomer conformation based on CyP-A The ability of change.Briefly, by by the peptide substrates supply response mixture including nitroaniline part, the reactant mixture Contain CYP-A, test compound (CsA analogs, CsA or dimethyl sulfoxide solvent) and second enzyme Chymetin.Each Test compound is in 10 concentration with triplicate or quadruplicate tested.Pass through on-catalytic and CYP catalysis process, the peptide Anti conformation is changed into from cisoid conformation.The transisomer of peptide, is not its cis-isomer, is the substrate of Chymetin. Chymetin cracks nitroaniline from remaining peptide immediately, and free nitroaniline is proportional with the ratio of cis-trans isomerization Accumulation.Because free nitroaniline is coloured product, it is accumulated, and by spectrophotometer measurement, its trap is quantified. The accumulation of paranitroanilinum is measured 6 minutes, and the first order rate constant of each reaction is calculated using Graphpad Prism softwares. By subtracting on-catalytic speed constant from overall reaction rate constant, (come from does not have the speed constant of each reaction of CyP-A catalysis CyP-A reaction) determined.Catalytic rate constant is demonstrated by its IC as the curve of inhibitor concentration function₅₀Value definition Compound potencies.

Detailed scheme

A. peptide

Measure is N- succinyl-alanines-Ala-Pro-phenylalanine-paranitroanilinum with peptide.By it with 3 MM concentration is dissolved in the solution of trifluoroethanol amine and lithium chloride (TFE/LiCl).By with 17 mg/ml concentration by lithium chloride Trifluoroethanol amine is dissolved in, fresh TFE/LiCl is made daily.After LiCl dissolvings, by the molecular sieve and gently for adding heated drying Mixed solution at least 30 minutes, reduces the water content in TFE/LiCl solution.Then peptide is dissolved in TFE/LiCl, it is cold before determining But solution is to 4 DEG C~8 DEG C.It is each determine reaction and start when, anhydrous TFE/LiCl peptide dissolution promote more polypeptide exist it is cis Conformation.Data analysis shows, about 60% peptide starts as cis-isomer in our measure, this and scientific literature report Data are consistent.In enzyme reaction, peptide is diluted 20 times to the final concentration that determines for 150 μ Μ.

B. test compound

Test compound is CsA, CsA analog or dimethyl sulfoxide (DMSO) (DMSO).By in sterile eppendorf tubes, Concentration is dissolved in DMSO for 10 mg/ml, the storing solution of CsA and CsA analogs is made.Storing solution is preserved when not in use At -20 DEG C.In the daily of measure, the test compound further diluted is made.In each experiment, respectively as solvent pair According to and reference compound, test DMSO and CsA.In microcentrifugal tube, the molecular weight based on compound is by CsA and CsA classes 50 μ Μ are diluted to DMSO like 10 mg/ml storing solutions of thing.Then in 96- hole polystyrene plates, it is made in DMSO and respectively changes Nine kinds of 3 times are serially diluted of compound.Aliquot DMSO- solution or single DMSO solvents (are seen below in reaction buffer Text formula) in dilution 50 times, be made CsA or CSA analogs final concentration of 1000,333,111,37,12,4.1,1.4, 0.46th, 0.15 and 0.05 nM.Before measure, the solution of reaction buffer is stored in 4 DEG C~8 DEG C at least one hour.

C. reaction buffer

The starting soln (salt buffer) of reaction buffer includes the mM of Hepes 50, the mM of sodium chloride 100 and human seralbumin The mg/ml of albumen 1, pH value is adjusted to 8.0 with sodium hydroxide.Salt buffer is stored in 4 DEG C when not in use.Each day is being determined, will Ox Chymetin is dissolved in the salt buffer of certain volume, and it is 1 mg/ml to make its concentration.By the Chymetin of aliquot Solution is removed, the reaction buffer compareed as on-catalytic.Into the Chymetin solution of remainder, people's restructuring is added CYP-A to concentration be 5 nM.Solution containing Chymetin and CYP-A is designated as reaction buffer, and anti-for preparing Answer liquid.

D. reaction scheme

It is all to determine reaction and carried out in the cold house of 4 DEG C~8 DEG C of temperature.Before the assay, all solution and equipment are in cold house Middle storage at least 1 hour.In order to carry out the measurement of available devices with speed slow enough, low temperature is that reaction is required.Measurement Device is the BMG Polarstar ELIASAs equipped with the nm trap readings of OD 405.Reaction is surveyed in 96- holes flat-bottomed polystyrene Carried out in fixed board.It is each determine operation by the row of plate one 12 independent reactions constitute.Peptide is moved with single channel in the row of plate one Then plate is placed under ELIASA plate supporter by liquid device per the μ l deciles of hole 5.95 μ l are reacted by using 12 channel pipettors Buffer solution is assigned to each hole containing peptide, and is thoroughly mixed each reaction by liquid relief repeatedly to ensure peptide uniform dissolution, starts anti- Should.Operating 12 reactions are respectively determined to be expressed as follows：

A) 10 reactions that a kind of 10 kinds of concentration of test compound are each repeated are represented (reaction buffer contains CyP-A)

B) 1 reaction of 5 μ l DMSO solvents (reaction buffer contains CyP-A)

C) 1 reaction of 5 μ l DMSO solvents (reaction buffer is free of CyP-A)

Trap record is immediately begun to upon mixing.Due to incorporation time and set instrument, from add reaction buffer to OD₄₀₅Record passes through about 15 seconds first.Follow-up reading was that interval has carried out 60 readings altogether for 360 seconds with 6 seconds.Each test Compound has carried out three or four reaction operations, to provide the data of parallel determination.

E. data analysis

Initial data is included in OD₄₀₅Time dependence increase.In the case where CYP-A exists and lacked with inhibitor, by OD₄₀₅'s Plateau proves that peptide was converted completely into transisomer in about 150 seconds., will using Graphpad Prism softwares OD₄₀₅Time data is marked and drawed, and is fitted to derive the first order rate constant K of each reaction with single-phase exponential equation.Do not having Have in CyP-A reaction, the spontaneity of the complete representative peptide of speed constant it is non-catalytic, hot it is cis-to-trans isomerization, It is defined as on-catalytic speed constant K₀.In the reaction containing CYP-A, isomerization is sent out by on-catalytic and enzymatic process It is raw.Therefore, on-catalytic speed constant K is represented in the speed constant K reacted containing CYP-A₀With catalytic rate constant K_catIt is total With.By subtracting K from total speed constant K₀(being obtained by the reaction without CyP-A) calculates K_cat.In 5 nM CyP-A, 150 μ In Μ peptide substrates and no inhibitor reaction, K_catUsual 3 times are higher than K₀。

By K_catTo inhibitor concentration drawing S-shaped dose response nonlinear regression and fitting, to prove inhibitor effectiveness.It is soft The EC that part is calculated₅₀Value represents test compound and suppresses 50%K_catConcentration.In order to which the variability between being tested under condition determination is entered Row standardization, CsA is run in each experiment as reference compound, and CsA analogs effect is based on EC₅₀Value is expressed as relatively CsA effect multiple.For example, CsA analogs EC₅₀It is CsA¹/₂, represent 2 times of the effect compared with CsA, and CSA analogs IC₅₀It is 5 times and is higher than CsA, represents 0.2 times compared with CsA of effect.

In table 5 shown in accompanying drawing, it is shown that in position 1 and the CsA analogs changed in position 1 and 3 according to the present invention Cyclophilin A suppress and immunosupress.

Claims

1. formula L compound：

Wherein

A. R' is H or acetyl group；

C. R2 is selected from：

I. unsubstituted, N- substitutions or N, N- disubstituded amide；

Ii. the amine that the substituted or unsubstituted acyl groups of N- are protected；

Iii. carboxylic acid；With

Iv. nitrile；With

D. R23 be methyl or ethyl,

Wherein described compound is D- epimers, wherein the chiral centre of the D- epimers is connected with R23 Carbon.

2. the compound of claim 1, wherein R1-R2 are selected from：

i. ；

ii. ；

iii. ；

iv. ；

v. ；

vi. ；

vii. ；

viii. ；

ix. ；

x. ；

xi. ；

xii. ；

xiii. ；

xiv. ；

xv. ；

xvi. ；

xvii. ；

xviii. ；

xix. ；

xx. ；

xxi. ；

xxii. ；

xxiii. ；

xxiv. ；

xxv. ；

xxvi. ；

xxvii. ；

xxviii. ；

xxix. ；

xxx. ；

xxxi. ；

xxxii. ；

xxxiii. ；

xxxiv. ；

xxxv. ；With

xxxvi. 。

3. the compound of claim 1, wherein R2 are selected from：

a. ；

b. ；

c. ；

d. ；With

e. ；

Wherein R5 is the saturation of 1 to 10 carbon lengths or the aliphatic carbon chain of undersaturated straight or branched.

4. the fat of the saturation of the compound of claim 1, wherein R1-R2 comprising 2 to 5 carbon or undersaturated straight or branched Race's carbochain, it is selected from the substituent substitution of the amine and carboxylic acid of the substituted or unsubstituted acyl group protection of nitrile, N-.

5. any one of claim 1-4 compound, wherein

Selected from following：

；

；With

。

6. the compound of claim 1 or claim 5, wherein R23 are ethyls.

7. the compound of claim 1 or claim 5, wherein R23 are methyl.

8. the compound of claim 7, wherein R1 are undersaturated C4-C15 straight-chain aliphatics carbochain and R2 is COOH.

9. the compound of claim 8, wherein R1 are undersaturated C6-C15 straight-chain aliphatics carbochains.

10. the compound of below general formula

Wherein：

R' is H or acetyl group；

R1-R2 is；With

R23 is ethyl or methyl, and

11. the compound of claim 10, wherein R23 are methyl.

12. the compound of below general formula

Wherein：

R' is H or acetyl group；

R1-R2 is；With

R23 is methyl, and

13. the method for the compound of claim 1-12 any one is prepared,

It comprises the following steps：

1) cyclosporin A (CsA) and alkaline alkyl lithium amide are reacted in the presence of suitable solvent, then with properly Electrophilic reagent react with the compound of production 1：

；

Formula 2A

3) formula 2A compound and oxidant are reacted, to form formula 3A compound：

Formula 3A

Formula 4A

5) formula 4A compound is carried out deacetylated.

14. the method for claim 13, wherein the preparation is included in step 1）In add in a suitable solvent it is excessive LiCl is to primarily form the L- epimers of compound, or the preparation is in step 1）In in a suitable solvent LiCl lack Lower progress is lost, to primarily form the D- epimers or L- epimerisms of the D- epimers of compound, wherein compound The chiral centre of body is the carbon atom being connected with R23.

15. the method for claim 13-14 any one, wherein the alkaline alkyl lithium amide is lithium diisopropylamine.

16. the method for claim 13-15 any one, wherein step 1) in the electrophilic reagent that uses it is fixed in following table The group of justice, to generate the corresponding R23 in table：

。

17. preparing the method for the compound of claim 1, it comprises the following steps：

Formula 1A compound is reacted with dimethylamino naphthyridine in the presence of appropriate solvent, to form the compound of formula 2：

Optionally subsequently form the aldehyde of formula 3：

Optionally then by Wittig reaction with production L compound, wherein R23 is methyl or ethyl.

18. preparing the method for the compound of claim 1, it comprises the following steps：Wherein R1 and R2 such as claim 1-4 are appointed The compound of formula 5 and alkaline alkyl lithium amide defined in what one, in the presence of suitable electrophilic reagent, in appropriate solvent Middle to be reacted, to form the compound of claim 1, wherein R23 is methyl or ethyl

。

19. the method for claim 18, wherein the alkaline alkyl lithium amide is lithium diisopropylamine.

20. the compound of claim 1-12 any one, wherein R' is H.

21. pharmaceutical composition, it includes the compound and one or more acceptable excipients of claim 1-12 and 20 any one Agent.